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1.
猪苓、伴生菌及蜜环菌共培养的形态学研究   总被引:4,自引:0,他引:4  
邢晓科  郭顺星 《菌物系统》2003,22(4):653-660
本文对猪苓、伴生菌及蜜环菌两两共培养及三者共培养进行了宏观形态观察及细胞学水平上的研究。结果表明,猪苓与伴生菌共培养时,在二者之间形成一致密拮抗线,猪苓菌落表面菌丝分化产生大量菌丝束;猪苓与蜜环菌共培养时,猪苓能阻止蜜环菌菌索对其自身的进一步侵袭,互作区中的双方菌丝及菌索均停止生长;蜜环菌与伴生菌共培养时,蜜环菌能穿透整个伴生菌菌落,在伴生菌菌落下方产生大量分枝;三者共培养后,猪苓对蜜环菌的防御能力有所下降,伴生菌对蜜环菌的耐受力有所提高,蜜环菌产生的新分枝均向伴生菌一侧生长,猪苓与伴生菌之间并不形成致密拮抗线,只可见双方菌丝的白色交融区。猪苓与伴生菌均能在蜜环茵菌索皮层上形成侵入位点。  相似文献   

2.
动物细胞的灌注培养方法   总被引:10,自引:0,他引:10  
随着基因技术的发展,细胞融合技术和杂交瘤技术在细胞中的应用,IFN、EGF、t-pA、UK、EPO等一些高价值的蛋白,相继在动物细胞中表达成功。但是,同传统的微生物细胞培养相比,动物细胞培养存在着细胞倍增时间长、代谢途径复杂、对营养的要求高、对外界环...  相似文献   

3.
应用免疫金标记技术证明,在眼虫藻和其它藻类中RuBP羧化酶主要分布在蛋白核部位,这与高等植物中RuBP羧化酶分布不同,在眼虫藻叶绿体间质中有少量RuBP羧化酶存在,这与高等植物中RuBP羧化酶的分布也有相似之处。暗中培养的眼虫藻不能形成类囊体,无RuBP羧化酶,无光合能力,只能进行异养代谢。  相似文献   

4.
邢晓科  郭顺星 《菌物学报》2003,22(4):653-660
本文对猪苓、伴生菌及蜜环菌两两共培养及三者共培养进行了宏观形态观察及细胞学水平上的研究。结果表明,猪苓与伴生菌共培养时,在二者之间形成一致密拮抗线,猪苓菌落表面菌丝分化产生大量菌丝束;猪苓与蜜环菌共培养时,猪苓能阻止蜜环菌菌索对其自身的进一步侵袭,互作区中的双方菌丝及菌索均停止生长;蜜环菌与伴生菌共培养时,蜜环菌能穿透整个伴生菌菌落,在伴生菌菌落下方产生大量分枝;三者共培养后,猪苓对蜜环菌的防御能力有所下降,伴生菌对蜜环菌的耐受力有所提高,蜜环菌产生的新分枝均向伴生菌一侧生长,猪苓与伴生菌之间并不形成致密拮抗线,只可见双方菌丝的白色交融区。 猪苓与伴生菌均能在蜜环菌菌索皮层上形成侵入位点。  相似文献   

5.
单细胞绿藻的大量培养试验   总被引:9,自引:0,他引:9  
在近廿年来,单细胞绿藻的培养和利用,曾引起广泛的兴趣,主要的是由于单细胞绿藻具有下列几个重要的特性:(1)能够充分利用太阳的光能,通过光合作用,制造大量的有机物质,作为动物的直接或间接的食料;(2)生长繁殖迅速,对自然环境的适应性很强,易于培养;(3)蛋白质的含量很高,  相似文献   

6.
植物组织培养的简化   总被引:16,自引:0,他引:16  
介绍简化植物组织培养中培养基、培养条件、培养器皿以及其他方面的简化所采取的方法和所取得的成果,并提出一套合理的简化植物组织培养、降低培养成本的方案。  相似文献   

7.
改善植物大规模组织培养条件的研究进展   总被引:11,自引:1,他引:10  
蔡能  易自力  李祥 《植物学通报》2003,20(6):745-751
植物组织培养技术具有巨大的应用价值,传统组织培养存在许多缺点,许多学者从培养基、培养容器、环境条件(光照、气体等)进行了研究和改善,取得了一定的成效。综述了这些方面的研究进展。  相似文献   

8.
本文对猪苓、伴生菌及蜜环菌两两共培养及三者共培养进行了宏观形态观察及细胞学水平上的研究。结果表明,猪苓与伴生菌共培养时,在二者之间形成一致密拮抗线,猪苓菌落表面菌丝分化产生大量菌丝束;猪苓与蜜环菌共培养时,猪苓能阻止蜜环菌菌索对其自身的进一步侵袭,互作区中的双方菌丝及菌索均停止生长;蜜环菌与伴生菌共培养时,蜜环菌能穿透整个伴生菌菌落,在伴生菌菌落下方产生大量分枝;三者共培养后,猪苓对蜜环菌的防御能力有所下降,伴生菌对蜜环菌的耐受力有所提高,蜜环菌产生的新分枝均向伴生菌一侧生长,猪苓与伴生菌之间并不形成致密拮抗线,只可见双方菌丝的白色交融区。 猪苓与伴生菌均能在蜜环菌菌索皮层上形成侵入位点。  相似文献   

9.
真菌深层培养过程的房室结构神经网络模型   总被引:1,自引:0,他引:1  
在对横纹黑蛋巢菌深层培养过程进行分析的基础上,提出一种房室结构的神经网络模型,利用RBF网络这各房室的输入,输出关系,并进一步对整个生化过程作了建模型研究,计算结果表明,所建模型性能较佳,对真菌培养过程的观测数据拟合结果令人满意。  相似文献   

10.
未培养的微生物研究进展   总被引:4,自引:0,他引:4  
未培养的微生物是指那些到目前为止人们还无法纯培养的微生物。这些微生物可能是以前被描述过的,更多的则是从未被了解过的。近年来,对未培养的微生物的研究方兴未艾,主要是有关环境微生物的遗传多样性分析,从特定环境中鉴定某些未知的微生物,从环境微生物中直接克隆基因以及提高环境微生物的培养率等问题。就有关这几个方面的一些研究进展作一个简要的介绍。  相似文献   

11.
The present study sought to determine whether water deprivation increases Fos immunoreactivity, a neuronal marker related to synaptic activation, in sympathetic-regulatory neurons of the hypothalamic paraventricular nucleus (PVN). Fluorogold (4%, 50 nl) and cholera toxin subunit B (0.25%, 20-30 nl) were microinjected into the spinal cord (T1-T3) and rostral ventrolateral medulla (RVLM), respectively. Rats were then deprived of water but not food for 48 h. Water deprivation significantly increased the number of Fos-positive nuclei throughout the dorsal, ventrolateral, and lateral parvocellular divisions of the PVN (water deprived, 215 +/- 23 cells; control, 45 +/- 7 cells, P < 0.01). Moreover, a significantly greater number of Fos-positive nuclei were localized in spinally projecting (11 +/- 3 vs. 2 +/- 1 cells, P < 0.025) and RVLM-projecting (45 +/- 7 vs. 7 +/- 1 cells, P < 0.025) neurons of the PVN in water-deprived vs. control rats, respectively. The majority of these double-labeled neurons was found in the ventrolateral and lateral parvocellular divisions of the ipsilateral PVN. Interestingly, a significantly greater percentage of RVLM-projecting PVN neurons were Fos positive compared with spinally projecting PVN neurons in the ventrolateral (25.8 +/- 0.7 vs. 8.0 +/- 1.5%, respectively, P < 0.01) and lateral (23.4 +/- 2.1 vs. 5.0 +/- 0.9%, respectively, P > 0.01) parvocellular divisions. In addition, we analyzed spinally projecting neurons of the RVLM and found a significantly greater percentage were Fos positive in water-deprived rats than in control rats (26 +/- 3 vs. 3 +/- 1%, respectively; P < 0.001). Collectively, the present findings indicate that water deprivation evokes a distinct cellular response in sympathetic-regulatory neurons of the PVN and RVLM.  相似文献   

12.
OBJECTIVE: To evaluate the patterns of testicular cytology in men with primary infertility, to compare the morphologic patterns between the periods 1990-1995, immediately after Gulf War II, and 1997-2001 and to determine whether there is a correlation between hormonal profile, testicular volume and morphologic pattern. STUDY DESIGN: Retrospective study of men with primary infertility. History, complete physical examination, hormonal assay and testicular ultrasound were evaluated. A total of 545 patients had samples for testicular cytology obtained from both testes. The patient's consent was obtained in all cases. Smears were interpreted under light microscopy after treatment with Diff-Quik. A total of 104 healthy, fertile subjects were used for comparison of the hormonal profile and testicular volume. RESULTS: The mean (+/- SD) age was 28.66 +/- 4.36 years and duration of marriage 4.4 +/- 4.36 years. There were 11.2% patients with normal cytology, 55.8% with hypospermatogenesis, 28.4% with testicular atrophy, 2.9% with Sertoli cells only and 1.7% with maturation arrest. A significant increase in hypospermatogenesis and decrease in the Sertoli cell-only pattern were noted in 1997-2001 when compared with 1990-1995. The mean left testicular volume was 10.53 +/- 5.51 mL3 in the infertile group vs. 15.2 +/- 4.97 in the fertile group (p < 0.003); right testicular volume was 10.84 +/- 4.77 vs. 15.15 +/- 5.31 (p < 0.003). The hormonal profile revealed higher luteinizing hormone and follicle-stimulating hormone levels in the infertile group vs. control group (8.53 +/- 9.03 and 16.44 +/- 19.243 vs. 6.98 +/- 4.53 and 7.37 +/- 6.63, respectively [p < 0.001]). Free testosterone was higher in the fertile group (39.69 +/- 12.76 vs. 20.28 +/- 8.5 [p < 0.000]). CONCLUSION: The majority of infertile males in our cohort had hypospermatogenesis; testicular atrophy was the next most common disorder. There was no major change in overall absolute numbers since the Gulf War. Testicular cytology by fine needle aspiration is a safe and well-tolerated complementary investigation for unexplained male infertility.  相似文献   

13.
During establishment of spermatogenesis at the prepubertal age, an early germ cells apoptotic wave occurs likely aimed to remove abnormal germ cells and to maintain a proper cell number ratio between maturating germ cells and Sertoli cells. Here we assessed Sertoli and germ cell apoptosis in relation to morphological parameters of Sertoli cell maturation in neonatal rats under the influence of testosterone, estradiol and FSH given alone or in combinations. From postnatal day (PND) 5th to 15th male rats were daily injected with: 1) 2.5 mg of testosterone propionate (TP), or 2) 12.5 microg of 17beta-estradiol benzoate (EB), or 3) TP+EB, or 4) 7.5 IU of human purified FSH (hFSH), or 5) hFSH+EB or solvents (control-C). Autopsy was performed on PND 16th. Sertoli cell nuclei area and incidence of seminiferous tubule lumen formation (LF) were taken as markers of Sertoli cell maturation. Sertoli and germ cell apoptosis was assessed using TUNEL method. In comparison with C, the area of Sertoli cell nuclei was significantly reduced after EB (25.7+/-2.0 vs. 30.9+/-1.6 microm2 for C, p<0.001) and increased after hFSH+EB (33.1+/-2.3 microm2, p<0.05). Incidence of LF was completely arrested by steroid hormone treatments given separately, significantly inhibited after TP+EB (median: 0.0%, vs. 2.0% for C p<0.05) and significantly enhanced after hFSH+EB (median: 51.0%, p<0.001). hFSH alone did not influence LF. Incidence of TUNEL positive Sertoli cells significantly increased after EB (median: 2.9% vs. 0.5% for C, p<0.05) or TP+EB (median: 2.2%, p<0.01) and was not affected by other treatments. Incidence of TUNEL positive germ cells increased significantly after EB alone (median: 4.4% vs. 2.5%, for C, p<0.01 ) and was significantly decreased by hFSH+EB (median: 0.5%, p<0.01). CONCLUSIONS: 1) Administration of testosterone or estradiol to immature rats inhibits Sertoli cell maturation. 2) Estradiol stimulates Sertoli and germ cell apoptosis while testosterone has no effect. 3) Testosterone eliminates estradiol--induced germ cell apoptosis when both hormones act in concert. 4) FSH in concert with estradiol, but neither one of the hormone alone, accelerate Sertoli cell differentiation and effectively inhibit germ cell apoptosis. 5) During seminiferous tubule maturation testosterone and the synergistic action of FSH with estradiol support germ cell survival while estradiol alone has an inhibitory, pro-apoptotic effect.  相似文献   

14.
Testicular descent was prevented unilaterally in newborn rats by cutting the gubernaculum testis. At 100 days of age, the number of Leydig and Sertoli cells per testis, the concentration of receptors for LH, FSH, prolactin and GnRH, and endogenous concentrations of progesterone and testosterone were determined. The weight of the abdominal testes was reduced by 80%, but in spite of this they contained as many Sertoli (32.8 +/- 1.3 X 10(6), mean +/- s.e.m., n = 6) and Leydig (28.2 +/- 1.7 X 10(6) cells as did scrotal testes (32.1 +/- 2.5 X 10(6) and 24.3 +/- 1.2 X 10(6) respectively). The numbers of receptors for LH (3.2 +/- 0.2 and 1.0 +/- 0.2 pmol/testis, mean +/- s.e.m., n = 11), FSH (358 +/- 11.0 and 96.3 +/- 12.6 fmol/testis) and prolactin (535 +/- 32.7 and 92.4 +/- 13.2 fmol/testis) were reduced (P less than 0.001) in abdominal testes, but the number of GnRH receptors was unaffected (8.9 +/- 1.4 and 12.1 +/- 1.8 fmol/testis, n = 6). Testicular testosterone concentration (30.9 +/- 4.4 vs 15.4 +/- 3.2 ng/g, n = 11, P less than 0.001), but not that of progesterone (0.87 +/- 0.10 vs 1.01 +/- 0.21 ng/g), was decreased in abdominal testes. The decreased receptor and androgen values reflect functional disturbances in the abdominal testes. The changed local milieu within abdominal testes may reduce hormone receptor concentrations which are then involved in the observed Leydig cell dysfunction.  相似文献   

15.
Much of what is known about the molecular regulation and function of adult Sertoli cells has been inferred from in vitro studies of immature Sertoli cells. However, adult and immature cells differ in significant ways and, moreover, many Sertoli cell functions are regulated by conditions that are difficult to replicate in vitro. Our objective was to develop a procedure to isolate Sertoli cells rapidly and in sufficient number and purity to make it possible to assess Sertoli cell function immediately after the isolation of the cells. The isolation procedure described herein takes less than 4 h and does not require culturing the cells. From a single 4-mo-old adult rat, we routinely obtain 7.0 +/- 0.4 x 10(6) Sertoli cells per testis, and from a 21-mo-old rat, 7.2 +/- 0.4 x 10(6) Sertoli cells per testis. The purity, determined by morphologic analyses of plastic-embedded cells or after staining for tyrosine-tubulin or vimentin, averaged 80%. The contaminants typically included germ cells (10%) and myoid cells (10%). The germ cell-expressed genes protamine-2 and hemiferrin were not detected in the Sertoli cell preparations by Northern blot analyses, but the Sertoli cell-expressed genes clusterin, cathepsin L, and transferrin were highly expressed. Transferrin mRNA levels were greater in Sertoli cells isolated from aged than from young adult rats, consistent with previous analyses of whole testes; and cathepsin L mRNA levels were far more highly expressed in Sertoli cells isolated from stages VI-VII than from other stages of the cycle of the seminiferous epithelium, also consistent with previous analyses of whole testes and isolated tubules. These studies indicate that the freshly isolated cells retain differentiated function, and thus it should be possible to assess the in vivo function of adult Sertoli cells by isolating the Sertoli cells and immediately assessing their function.  相似文献   

16.
Stereological study of postnatal testicular development in Blackbelly sheep   总被引:2,自引:0,他引:2  
The objective was to characterize testicular development in Blackbelly sheep, focusing primarily on Sertoli cell number. Lambs (n=43) were allotted into eight groups, and surgically castrated at 0, 3, 6, 9, 12, 15,18 or 21 weeks of age (n=4-6 lambs per group). Testes were fixed and paraffin-embedded, cross-sections (5 microm) were stained and evaluated with quantitative morphometry techniques. Testis weight increased at a greater rate between 9 and 15 weeks of age, which was associated with remarkable changes in testicular histology, including increases in tubular tissue volume, and tubule diameter and length. Spermatogenesis started in a period between 9 and 12 weeks, lumen and elongated spermatids were observed for the first time at 12 weeks (78% of the tubules) and 15 weeks (37% of the tubules), respectively. Total number of Sertoli cells (mean+/-S.E.M.) increased steadily from birth (531+/-76 x 10(6)) to 15 weeks (12,008+/-1722 x 10(6)), with no changes afterwards. Sertoli cell number per gram of testicular tissue decreased as lambs were older, with the most remarkable change occurring between Weeks 9 and 12. An early increase in serum LH was observed at 6 weeks of age, with testosterone (T) increasing at Weeks 12 and 21. In conclusion, Sertoli cells maintained the capacity of proliferating from birth to 15 weeks of age in Blackbelly sheep; furthermore, the period of accelerated testis growth was associated with increased serum T concentration and with important changes in testicular morphology, as a consequence of the beginning and establishment of spermatogenesis and Sertoli cell maturation.  相似文献   

17.
Our purpose was to evaluate cellular androgen receptor (AR) distribution and intensity of immunostaining in the human azoospermic testis. Thirty six biopsy specimens from azoospermic men were immunostained, using a monoclonal antibody of human AR. The localization and the intensity of AR immunostaining was evaluated in Sertoli Cell Only (SCO) testis (G1, n = 21), in spermatogenesis arrest testis (G2, n = 11) and in histologically normal testis (G3, n = 4). We found an AR immunostaining in Sertoli, peritubular myoid and Leydig cells, but not in germ cells. The intensity of the immunostaining varied substantially between biopsy specimens of different patients. Sertoli and Leydig cells AR immunostaining (score and intensity) in SCO group was higher than in the other groups. For Sertoli cells, the score means of AR immunoreactivity were 20 +/- 2.36, 10.18 +/- 1.0 and 1 +/- 1, for G1, G2 and G3 groups, respectively. For Leydig cells, the score means were 10.24 +/- 1.37, 6 +/- 0.71 and 0, for G1, G2 and G3 groups, respectively. We found significant differences between G1 and G2 (p = 0.0008), between G1 and G3 (p = 1.54 10-7) and G2 and G3 (p = 0.00032). These results suggest that in the testis AR is located exclusively in somatic cells and its expression is higher in SCO syndrome than in normal and in arrest spermatogenesis testes.  相似文献   

18.
Yang X  Han JQ  Liu R 《生理学报》2008,60(1):143-148
本文旨在探讨肠道局部炎症对脊髓肠道感觉传入神经通路的近期及远期效应,应用三硝基苯磺酸(trinitrobenzenesulfonic acid,TNBS)建立大鼠结肠炎动物模型,用DiI(3)逆行神经标记法识别支配肠道炎症部位的脊髓背根神经节(dorsalrootganglia,DRG)神经元,通过肉眼观察、平均组织损伤评分及髓过氧化物酶活性测定等方法评价肠道组织的炎症反应状态,用免疫组织化学法测定香草酸受体l(vanilloid receptor 1,VRl)和降钙素基因相关肽(calcitonin gene-related peptide,CGRP)在支配结肠炎症部位的DRG神经元中的表达,比较炎症不同阶段(给予TNBS后7、21、42 d)CGRP和VRI阳性神经元的数目.结果显示,炎症急性期(即给予TNBS后7 d)结肠黏膜肉眼可见明显损伤,同时相应DRG中表达CGRP及VRl的神经元增加近2倍[(95.38±9.45)%VS(42.86±5.02)%,(89.23±8.21)%VS(32.54±4.58)%].给予TNBS后21、42 d,肠道炎症反应已完全消退,但表达CGRP及VRl的DRG神经元数目仍明显高于对照组[(86.25±8.21)%,(68.28±7.12)%VS(42.86±5.02)%;(67.22±6.52)%,(56.25±4.86)%VS(32.54±4.58)%].结果提示,肠道局部炎症可以上调支配肠道的脊髓传入神经元中CGRP和VRl的表达,这种异常表达可以持续至肠道炎症反应消退后的一定时间.  相似文献   

19.
The aim of this study was to examine the effects of timolol in an experimental model of elevated intraocular pressure (IOP). Three episcleral veins of rats with normal IOP were cauterized. Three months later we examined the effects on anterograde axonal transport from the retinal ganglion cells (RGCs) to the superior colliculus (SC) as well as on the number of neurons in the retinal ganglion layer (RGL). These parameters were also studied in a group of rats submitted to treatment with timolol after confirming that their IOP was still raised after two weeks. After the surgical procedure, the mean IOP of the experimental eyes increased to 33.5+/-1.06 mmHg (1.25 fold compared to the control group) and three months later the IOP remained significantly elevated; however, after a long period of treatment with timolol the IOP was 14.05+/-0.81 mmHg, similar to that of the control group. In the group with normal IOP, labelling with horseradish rabbit peroxidase (HRP) at 120 minutes and 24 hours postinjection showed continuous staining from the retina to the SC. In the experimental group the optic nerve head (ONH) was completely negative, although in the group treated with timolol there was partial block of axonal transport in the ONH, in which the staining was slightly more intense. The number of neurons in the RGL, counted by immunohistochemical labelling with Neu-N, showed that in eyes with normal and elevated IOP there were 423+/-11 neurons/mm(2) and 283+/-10 neurons/mm(2), respectively. After treatment with timolol the number of neurons (331+/-10 cells/mm(2) increased compared with elevated IOP eyes, although the number did not reach that of the control group. These results indicate that treatment with timolol, started two weeks after the surgical procedure, was partially neuroprotective because the loss of neurons in the RGL was lower than in untreated animals, though not sufficient to re-establish normal axonal transport.  相似文献   

20.
Meng XT  Li C  Dong ZY  Liu JM  Li W  Liu Y  Xue H  Chen D 《Cell biology international》2008,32(12):1546-1558
We have previously demonstrated that amniotic epithelial cells (AECs) can enhance survival and neural differentiation of neural stem cells (NSCs) when co-cultured in basal media. In addition, the presence of basic fibroblast growth factor (bFGF) enhances this AEC function. The aim of the present study was to extend those findings and investigate whether AECs modified with the bFGF gene will also enhance NSCs survival and neural differentiation in vivo and promote repair of the injured spinal cord. Female Wistar rats were used for a contusive spinal cord injury (SCI) model. Contusive SCIs were induced using a weight-drop device at levels T9-T11. Seven days following contusion, rats received grafts of NSCs only, NSCs with AECs/pLEGFP-hbFGF, or NSCs with AECs/pLEGFP-C1 into the injured region. Significant locomotor improvement was observed in the NSCs/AECs co-graft group beginning at 3 weeks compared with the NSCs or NaCl only groups. These results were confirmed and extended in an electrophysiological analysis. An immunohistological analysis revealed that AECs/pLEGFP-hbFGF promoted the survival (vs NaCl group: 194+/-9.17 vs 103.6+/-13.05) and neural differentiation (vs NaCl group: 14.24+/-1.11 vs 7+/-0.63) of co-transplanted NSCs. We also confirmed that AECs could promote the survival of host neurons. These results suggest that AECs/pLEGFP-hbFGF improve the NSCs survival and differentiation microenvironment and may be useful as a source of sustained trophic supported to improve NSCs differentiation into neurons in vivo. These findings suggest that a cograft of AECs/pLEGFP-hbFGF and NSCs may have benefits for SCI.  相似文献   

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