黄芪注射液对日本血吸虫成虫细胞鸟氨酸脱羧酶活性作用的研究

目的:观察黄芪注射液作用后日本血吸虫成虫细胞鸟氨酸脱羧酶活性的变化。方法:细胞接种后1周时,分别用浓度为2μg/ml、5μg/ml和8μ/ml的黄芪注射液处理24小时,然后用RPMI-1640加10%小牛血清的培养基培养。用分光光度法测定细胞鸟氨酸脱羧酶活性。结果:浓度为2μ/ml的黄芪注射液处理后第4周时细胞鸟氨酸脱羧酶活性显著增高。结论:黄芪注射液有增强日本血吸虫成虫细胞鸟氨酸脱羧酶活性的作用。

PHA对日本血吸虫成虫细胞鸟氨酸脱羧酶活性作用的研究

目的：研究植物血球凝集素（ＰＨＡ）对日本血吸虫成虫细胞的促增殖作用。方法：观察ＰＨＡ对日本血吸虫成虫细胞鸟氨酸脱羧酶活性的影响。在日本血吸虫成虫细胞接种 １周后分别用浓度为 １００ ｇ／ｍｌ、２００ ｇ／ｍｌ、３００ ｇ／ｍｌ和 ４００ ｇ／ｍｌ的 ＰＨＡ处理 ２４小时，然后用 ＲＰＭＩ—１６４０加１０％小牛血清的培养基培养。用分光光度法测定细胞鸟氢酸脱羧酶活性。结果；浓度为 ２００ ｇ／ｍｌ的ＰＨＡ处理后第２、３周时的细胞鸟氨酸脱羧酶活性显著增高。结论：ＰＨＡ有促进日本血吸虫成虫细胞增殖的作用。

日本血吸虫成虫培养细胞SDH和LDH细胞化学研究

目的 研究日本血吸虫成虫培养细胞琥珀酸脱氢酶 (SDH)和乳酸脱氢酶 (LDH)含量、分布及变化规律 ,了解日本血吸虫成虫培养细胞的能量代谢类型。方法 将虫龄 2 6d的日本血吸虫成虫细胞接种于小盖玻片上 ,培养于RPMI- 16 4 0含2 0 %小牛血清附加常量抗生素的培养基中 ,定时运用Pearson法进行SDH和LDH染色 ,用Olympus-BH2 显微镜观察并拍照 ,用HPIAS - 2 0 0 0图像分析仪测量其含量 ,并作统计分析。结果 日本血吸虫成虫培养细胞均具有SDH和LDH活性 ,两者均分布在细胞质内。培养 1d细胞的SDH和LDH活性最强 ,随着培养时间延长 ,其活性逐渐减弱 ,其中SDH活性下降较快 ,培养 5d大部分细胞SDH活性已极弱 ;而LDH活性下降则较缓 ,培养 5 6d细胞仍具LDH活性。结论 体外培养的日本血吸虫成虫细胞的能量代谢类型与成虫相似 ,既存在三羧酸循环需氧型呼吸链 ,也具有无氧糖酵解 ,但以无氧糖酵解为主。

甲基硝基亚硝基胍对日本血吸虫成虫培养细胞骨架作用的研究

目的 通过观察 N-甲基 - N-硝基 - N-亚硝基胍 (MNNG)诱导日本血吸虫培养细胞后骨架的改变 ,研究 MNNG对培养细胞转化、增殖的作用。方法 将日本血吸虫成虫培养细胞接种于小盖玻片上 ,培养 1周时用浓度为 3mg/ L 的 MNNG处理 48h,然后用 RPMI- 16 40含 2 0 %小牛血清附加常量抗菌素的培养基培养 2周 ,再换用 RPMI- 16 40含 5 %小牛血清的培养基继续培养 ,用考马斯亮兰染色法和鬼笔环肽荧光染色法显示培养细胞骨架。结果 MNNG诱导后 ,在培养第 4、5周时部分培养细胞骨架排列紊乱 ,微丝集聚于细胞边缘。结论 MNNG有一定诱导日本血吸虫成虫培养细胞转化。

EGF对用或未用MNNG诱导的日本血吸虫成虫培养细胞生长的影响

目的 研究表皮生长因子 (EGF)对用或未用甲基硝基亚硝基胍 (MNNG)诱导的日本血吸虫成虫培养细胞的促生长、增殖作用。方法 将联合法接种培养至第 3天的日本血吸虫成虫培养细胞 ,一部分在含 EGF终浓度分别为 0、0 .5、1、2、4、8、12、16、2 0、2 4、2 8ng/ml的附加 2 0 %小牛血清及常量抗菌素的 RPMI- 16 40培养基中培养 ;另一部分 ,先用含 MNNG终浓度为 3μg/m l附加 2 0 %小牛血清和常量抗生素的 RPMI- 16 40培养基处理48h,再换用含终浓度分别为 0、1、8、12、16、2 0 ng/ml的 EGF培养基继续培养。每日用 Olym pus IM倒置显微镜观察细胞生长和增殖情况。结果 两实验组中 ,用不同浓度 EGF处理后的日本血吸虫成虫培养细胞均在 2周以后出现不同程度的脱落、退化 ;并且 ,随着 EGF浓度的升高 ,培养细胞脱落、退化的现象更加严重 ;两实验组相比 ,未用 MNNG诱导的培养细胞出现脱落和退化现象的时间比用 MNNG诱导的相对更早。结论 外加 EGF对用或未用 MNNG诱导的日本血吸虫成虫培养细胞的生长增殖均无促进作用 ,相反 ,加速了培养细胞的老化 ,尤其是对未用 MNNG诱导的日本血吸虫成虫培养细胞 。

日本血吸虫成虫培养细胞骨架系统的研究

目的 研究日本血吸虫成虫培养细胞的骨架系统。方法 将日本血吸虫成虫细胞接种于小盖玻片上 ,以 RPMI- 16 40含 2 0 %小牛血清附加常量抗菌素的培养基培养 ,用考马斯亮蓝染色法和鬼笔环肽荧光染色法显示培养细胞骨架。结果 考马斯亮蓝染色显示日本血吸虫成虫细胞骨架呈网络状结构 ,均匀分布于胞质中 ;用 (NH4) 2 SO4抽提后考马斯亮蓝染色显示细胞骨架无明显改变 ;鬼笔环肽荧光染色后可见细胞内有均匀荧光亮点。结论 日本血吸虫成虫培养细胞骨架呈致密网络状结构 ,以中间纤维为主 ,缺乏粗大的微丝束。

柞蚕成虫卵巢细胞株的建立及其对病毒敏感性的研究

建立了一株连续生长的柞蚕成虫卵巢细胞株Ap－4，测定了细胞株的生长曲线，细胞株的群体倍增时间为67h，该细胞株亦可以在无血清培养基SF－900Ⅱ中生长。用同源的柞蚕核型多角体病毒感染细胞后，观察了细胞的病理变化以及病毒多角体的形成。检测了病毒多角体的数量以及非包埋型病毒的TCID50数值，细胞株增殖的病毒对柞蚕幼虫和Ap－4细胞株仍具有感染性。

日本血吸虫细胞凋亡及其诱导的研究

[目的 ]探讨日本血吸虫的细胞凋亡现象及诱导日本血吸虫细胞凋亡的可能性。 [方法 ]分离、培养日本血吸虫成虫混合细胞和虫卵。用原位末端标记法和琼脂糖凝胶电泳法分别检测正常培养条件下和诱导因素存在时细胞凋亡发生情况。 [结果 ]未经诱导的成虫细胞 ,DNA琼脂糖凝胶电泳未见凋亡所特有的“梯形”条带 ,成虫细胞核可见少量明显的凋亡所特有阳性着色。经诱导的成虫细胞和卵细胞琼脂糖凝胶电泳均呈凋亡所特有的“梯形”条带 ,诱导组的成虫细胞核可见明显的凋亡所特有的阳性着色和凋亡小体。 [结论 ]首次观察到日本血吸虫成虫细胞存在凋亡现象 ,且外源性因素可诱发日本血吸虫成虫细胞和卵细胞凋亡。

大戟对血吸虫细胞鸟氨酸脱羧酶活性的作用

目的 通过观察大戟水提取液对日本血吸虫成虫细胞鸟氨酸脱羧酶活性的影响 ,研究其对该种细胞转化、增殖的作用。方法 在细胞接种 1周后分别用浓度为 10、2 0、30、4 0 g/L 的大戟水提取液处理 2 4 h,然后用 RPMI- 16 4 0加 10 %小牛血清的培养基培养。用分光光度法测定细胞鸟氨酸脱羧酶活性。结果 浓度为 30 g/L的大戟水提取液处理后第 2、3周时细胞鸟氨酸脱羧酶活性显著增高。

The ORF 113 of Heliocoverpa armigera Single Nucleopolyhedrovirus Encodes a Functional Fibroblast Growth Factor

Fibroblast growth factor (FGF) is a key regulator of developmental processes. A FGF homolog (vFGF) is found in all lepidopteran baculoviruses. Autographa californica nucleopolyhedrovirus (AcMNPV) and Bombyx mori NPV (BmNPV) vFGFs are chemotactic factors. Here we analyzed the vfgf of Helicoverpa armigera NPV (HearNPV), a group II NPV. The HearNPV vfgf transcripts were detected from 18 to 96 h post-infection (hpi) of Hz-AM1 cells with HearNPV and encoded a 36 kDa protein, which was secreted into the culture medium. HearNPV vFGF had strong affinity to heparin, a property important for FGF signaling via an FGF receptor. Unlike its AcMNPV homolog, HearNPV vFGF specially chemoattracted Hz-AM1, but not other insect cells such as Sf9 and Se-UCR and not the mammalian cells 293 and HepG2. HearNPV vFGF is also associated with the envelope of BV but is absent in occlusion-derived virus, which coordinated to the chemotatic activity analysis.

Polysaccharides Purified from Wild Cordyceps Activate FGF2/FGFR1c Signaling

Land animals as well as all organisms in ocean synthesize sulfated polysaccharides. Fungi split from animals about 1.5 billion years ago. As fungi make the evolutionary journey from ocean to land, the biggest changes in their living environment may be a sharp decrease in salt concentration. It is established that sulfated polysaccharides interact with hundreds of signaling molecules and facilitate many signaling transduction pathways, including fibroblast growth factor （FGF） and FGF receptor signaling pathway. The disappearance of sulfated polysaccharides in fimgi and plants on land might indicate that polysaccharides without sulfation might be sufficient in facilitating protein ligand/receptor interactions in low salinity land. Recently, it was reported that plants on land start to synthesize sulfated polysaccharides in high salt environment, suggesting that fungi might be able to do the same when ex- posed in such environment. Interestingly, Cordyceps, a fungus habituating inside caterpillar body, is the most valued traditional Chi- nese Medicine. One of the important pharmaceutical active ingredients in Cordyceps is polysaccharides. Therefore, we hypothesize that the salty environment inside caterpillar body might allow the fungi to synthesize sulfated polysaccharides. To test the hypothesis, we isolated polysaccharides from both lava and sporophore of wild Cordyceps and also from Cordyceps militaris cultured without or with added salts. We then measured the polysaccharide activity using a FGF2/FGFRlc signaling-dependent BaF3 cell proliferation assay and found that polysaccharides isolated from wild Cordyceps activated FGF2/FGFR signaling, indicating that the polysaccha- rides synthesized by wild Cordyceps are indeed different from those by the cultured mycelium.

Microtubular elements of cortex in Euplotes Woodruffi

The cortical ciliature microtubular organelles of the ciliated protozoan Euplotes Woodruffi were analyzed with FLUTAX and anti-a-tubulin antibody. It showed that the cortical cytoskeleton was composed of non-ciliature structure, ciliature structure including adoral zone of membranelles （AZM）, undulating membranes （UM）, frontal-ventral-transverse cirri （FVTC）, caudal cirri （CC）, dorsal kineties, as well as base-associated microtubules. The silverline system is composed of longitudinal and transverse microtubules, concave structure appeared on the cell dorsal side, the base of DK containing rosette-like skeleton structure. All these suggested that the non-ciliature structure, ciliature structure of Euplotes be quite different from that of other species of ciliate, the silverline-system be true pare of cortical microtubute cytoskeleton in Euplotes, rosette structure be part of the basecytoskeleton.

Antigenicity of Synthetic Peptides Derived from Plasmodium Apoptosis-Linked Pathogenicity Factors

Background： Plasmodium falciparum malaria remains a major life-threatening disease. Recently, the Plasmodium apoptosis-linked pathogenicity factors （PALPF） have been identified. These antigens PALPF are expressed only by P falciparum-infected erythrocytes triggering endothelial cell apoptosis （apoptogenic）. Methods： We designed ten synthetic peptides （PI to P10） from PALPF： PF07 0032, PF10_0226, PFI0130c, PFD0875c and MAL13P1.206, and analyzed their antigenicity with an ELISA method using plasma samples from subjects living in Dienga, Gabon. Results： Four peptides showed good reactivity with human antibodies. The prevalence rate of specific IgG was 61%, 51%, 44% and 34% for P5, P6, P4 and P2, respectively. The median optical density of total IgG anti-P2 was higher than that directed against P4 and P6 （P = 0.009; P = 0.012 respectively）. The prevalence rate oflgG subclasses determined with plasma samples recognizing peptide 5 for IgGl, 2, 3 and 4 isotypes was 69%, 45%, 76% and 62%, respectively. All the subjects had at least one immunoglobulin subclass, while 13 （44%） had both IgG1 and IgG3 antibodies. There was no significant difference in the prevalence rate of anti-P5 IgG1, IgG3 and IgG4. Conclusion： These results warrant further immunogenicity studies of peptides 2, 4, 5 and 6 with a view of a tentative to antimalarial vaccine development.

Giardia infection:Protein-losing enteropathy in an adult with immunodeficiency

The case of a 52-year-old woman with a past history of thymoma resection who presented with chronic diarrhea and generalized edema is the focal point of this article.A diagnosis of Giardia lamblia infection was established,which was complicated by protein-losing enteropathy and severely low serum protein level in a patient with no urinary protein loss and normal liver function.After anti-helmintic treatment,there was recovery from hypoalbuminemia,though immunoglobulins persisted at low serum levels leading to the hypothesis of an immune system disorder.Good's syndrome is a rare cause of immunodeficiency characterized by the association of hypogammaglobulinemia and thymoma.This primary immune disorder may be complicated by severe infectious diarrhea secondary to disabled humoral and cellular immune response.This is the first description in the literature of an adult patient with an immunodeficiency syndrome who presented with protein-losing enteropathy secondary to giardiasis.

Characterization of the viral fibroblast growth factor homolog of Helicoverpa armigera single nucleopolyhedrovirus

Fibroblast growth factor(FGF) is found throughout multicellular organisms; however, fgf homologs(vfgf) have only been identified among viruses in lepidopteran baculoviruses. The function of v FGFs from Group I alphabaculoviruses, including Autographa californica multiple nucleopolyhedrovirus(Ac MNPV) and Bombyx mori nucleopolyhedrovirus(Bm NPV), involves accelerated killing of infected larvae by both viruses. The v FGF of Group II alphabaculovirus is structurally different from that of Group I alphabaculovirus, with a larger C-terminal region and additional N-linked glycosylation sites. In this study, we characterized the Group II alphabaculovirus v FGF of Helicoverpa armigera single nucleopolyhedrovirus(Hear NPV). The transcription and expression of vfgf was detected at 3 h and 16 h post-infection in Hear NPV-infected cells. To further study v FGF function, we constructed vfgf-knockout and-repaired Hear NPV bacmids and investigated their affect in both cultured cells and insects. Deletion of vfgf had no effect on budded-virus production or viral DNA replication in cultured Hz AM1 cells. However, bioassays showed that Hear NPV vfgf deletion significantly increased the median lethal dose and delayed the median lethal time by ~12 h in the host insect when the virus was delivered orally. These results suggested that v FGF is an important virulent factor for HearN PV infection and propagation in vivo.