阴道毛滴虫染色体组型观察

采用火焰干燥法制备阴道毛滴虫染色体标本,按 Levan等方法,进行核型分析。阴道毛滴虫的染色体数目为n＝10,2n ＝20,染色体的组型为3m、3sm、1st和3T。 Abstract：　The chromosomes ofTrichomonas vaginaliswere prepared by flame-drying method. Data of chromosome were measured and calculated. According to Levan's method, analysis of karyotype was described. The results showed that the chromosome number ofT. vaginalisconsisted of twenty pairs(2n=20) and chromosome type was 3m, 3sm, 1st and 3T.     

阴道毛滴虫染色体组型观察

采用火焰干燥法制备阴道毛滴虫染色体标本,按 Levan等方法,进行核型分析。阴道毛滴虫的染色体数目为n＝10,2n ＝20,染色体的组型为3m、3sm、1st和3T。 Abstract：　The chromosomes ofTrichomonas vaginaliswere prepared by flame-drying method. Data of chromosome were measured and calculated. According to Levan's method, analysis of karyotype was described. The results showed that the chromosome number ofT. vaginalisconsisted of twenty pairs(2n=20) and chromosome type was 3m, 3sm, 1st and 3T.     

探讨荆芥体外抗阴道毛滴虫的效果

目的探讨荆芥体外抗阴道毛滴虫的效果。方法将不同浓度的荆芥水提液作用于体外培养的阴道毛滴虫,于药物作用后不同时问记录毛滴虫的死亡率,并在光镜下观察药物作用前后毛滴虫的形态变化,同时与白头翁和青蒿的体外杀虫效果相比较。结果荆芥的杀虫效果与药物浓度和作用时间呈正相关。荆芥水提液触杀阴道毛滴虫的最低有效浓度为1：4。3种中药中,白头翁的杀虫效果最好,与另2种中药相比较差异有非常显著性（P〈0．01）;荆芥与青蒿的作用效果接近（P〉0．05）。荆芥作用后毛滴虫体内充满大量颗粒和空泡,部分虫体裂解、内容物外溢。结论荆芥具有较强的抗阴道毛滴虫作用。     

荆芥体外抗阴道毛滴虫作用机制的实验研究

目的探讨荆芥体外杀灭阴道毛滴虫的作用机制。方法将浓度为1∶4的荆芥水提液作用于体外培养的阴道毛滴虫,并采用透射电镜观察经药物作用2 h和4 h后阴道毛滴虫的超微结构变化。结果荆芥作用后阴道毛滴虫多聚核糖体解聚,粗面内质网脱颗粒,胞质内可见大量空泡;随药物作用时间延长,核膜不完整,核质变疏松;最终,胞膜破损,内容物外溢,虫体死亡。结论荆芥可破坏阴道毛滴虫的内膜系统,具有较强的抗滴虫作用。     

5—硝基咪唑类药物体外抗无芽孢厌氧菌效果的观察

从多种临床标本中分离并鉴定了200株无芽孢厌氧菌.赛克硝唑(SNZ)、替硝唑(TNZ)和甲硝唑(MNZ)对120株革兰氏阴性厌氧菌的MIC90分别为1～4,1～2和4～8 mg/L,对80株革兰氏阳性厌氧菌的MIC90分别为4～8,4～16和16～32 mg/L.SNZ体外抑制革兰氏阴性厌氧菌作用略弱于TNZ,但抑制革兰氏阳性无厌氧菌较TNZ稍强,MNZ对两类厌氧菌的抑制效果均不如SNZ和TNZ.     

阴道毛滴虫铁氧还蛋白基因的原核表达

目的在原核细胞中表达阴道毛滴虫铁氧还蛋白(ferredoxin,Fd)基因。方法构建阴道毛滴虫Fd基因的原核表达重组质粒pUC19-Fd,转化大肠埃希菌JM109感受态细胞中,异丙基-β-D-硫代半乳糖苷(IPTG)诱导蛋白质表达。结果经十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹(Western blot)分析,重组质粒在大肠埃希菌中表达出Fd。结论在大肠埃希菌中表达出了Fd。     

阴道毛滴虫多克隆抗体的制备及Fd基因的原核表达

目的 将阴道毛滴虫铁氧还蛋白（ferredoxin,Fd）基因在大肠埃希菌中诱导表达。方法制备阴道毛滴虫可溶性抗原,多点注射免疫家兔,获得的血清用ELISA测定其抗体效价。将原核表达重组质粒pET3C-Fd转化入大肠埃希菌BL21（DE3）感受态细胞中,异丙基-β-D-硫代半乳糖苷（IPTG）诱导蛋白质表达。结果制备出抗阴道毛滴虫多克隆抗体,抗体效价在1：8000以上,用于免疫印迹实验。经十二烷基磺酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和免疫印迹（Western blot）分析,重组质粒在大肠埃希菌中表达出Fd。结论在大肠埃希菌中表达出了Fd。     

阴道卷曲乳酸杆菌对阴道致病菌的体外生物拮抗作用

目的研究阴道卷曲乳酸杆菌对阴道致病菌的体外生物拮抗作用。方法将阴道卷曲乳酸杆菌分别与3株阴道致病菌（金黄色葡萄球菌、大肠埃希菌、粪肠球菌）混合接种于GAM液体培养基中进行厌氧培养,通过平板菌落计数法计算阴道致病菌的菌数。结果经混合培养后阴道致病菌的菌数明显下降。结论阴道卷曲乳酸杆菌A7在体外能明显抑制3株肠道致病菌的生长繁殖。     

薄荷油体外抗蠕形螨效果及杀螨机制

采用透明胶带粘贴过夜法获取2种人体蠕形螨,随机分组,观察不同浓度的薄荷油的杀虫效果,并在MoticDMB5图像采集软件系统下拍摄虫体在薄荷油作用下的虫体死亡过程。结果表明,薄荷油有很强的体外杀灭2种人体蠕形螨的作用,尤其对皮脂蠕形螨的杀灭作用显著。随着药物浓度的增加及作用时间的延长,蠕形螨死亡率增高。12.5%、3.125%分别是薄荷油体外杀灭毛囊蠕形螨和皮脂蠕形螨的最适杀螨浓度。薄荷油作用于蠕形螨,可见虫体收缩扭动,活动加剧,消化管剧烈收缩,毛囊蠕形螨和皮脂蠕形螨体壁均有渗出物外溢。虫体表现为兴奋—痉挛—失水—松弛—死亡的典型症状。薄荷油对人体蠕形螨的杀灭机制主要通过神经毒性和直接毒杀作用,造成虫体破裂脱水而死亡。薄荷油是一种极具开发潜能的高效的天然杀螨药。     

小鼠卵母细胞体外成熟、体外受精的效果观察

目的　研究不同培养条件对小鼠卵母细胞体外成熟及体外受精率的影响。方法　小鼠卵母细胞分别在含有FSH、BSA和胰岛素的培养液中体外成熟,在Whitten 氏液中体外受精,比较体外成熟率、体外受精率。结果　1- 裸卵(DO) 的体外成熟率、体外受精率(81-4% ,31-0 % ) 均高于卵丘卵母细胞复合体(COC)(48-6 % ,27-1% ) 。2- 在培养液中添加FSH、胰岛素和BSA,卵母细胞的体外成熟率为77-9 % ,82-3% 、60-7% ;体外受精率为77-2 % 、72-6 % 、26-7% ;2 - 细胞率为49-2 % 、34-2 % 、10-0% 。胰岛素组的卵母细胞IVM 率最高,但IVF率、2 - 细胞率低于FSH 组。3- 添加BSA的两组的体外受精率只有26-7 % 、25-8 % ,显著低于其他组,其体外成熟率也较添加FSH 和胰岛素的组成。4- 排出第一极体(PbI) 的卵母细胞的体外受精率和2 - 细胞率(85-9 % ,22-4% ) 均高于GV期卵母细胞(71-1 % ,12-9 % ) 。结论　1- 卵丘卵母细胞(COC) 较裸卵(DO) 的体外成熟率、体外受精率都低,差异显著(P成熟< 0-01;P受精< 0-05) 。2-FSH 和胰岛素均能提高小鼠卵母细胞的体外成熟率、体外受精率。3-BSA可以降低小鼠卵母细胞体外受精率,差异极显著。4-GV 期卵母细胞的体外受精率显著低于体外培养的排出第一极体的卵母细胞(P2 - cell < 0-05,P受精<0-05)     

Trichomonas vaginalis: observation of coexistence of multiple viruses in the same isolate

Trichomonas vaginalis is a flagellated, parasitic protozoan that inhabits the urogenital tract of humans. Some isolates of T. vaginalis are infected with a double-stranded RNA (dsRNA) virus, which was described in the literature as homogeneous icosahedral viral particles with an isometric symmetry and 33 nm in diameter. This study examined in detail the viral particles in T. vaginalis isolate 347 and describes a heterogeneous population of viral particles. The different dsRNA viruses were only observed after a change in the technique. The sample was prepared by the negative staining carbon-film method directly onto freshly cleft mica. The detected viruses ranged in size from 33 to 200 nm. Among the shapes observed were filamentous, cylindrical, and spherical particles. These results show that T. vaginalis may be a reservoir for several different dsRNA viruses simultaneously.     

Phagocytosis by Trichomonas vaginalis: new insights

BACKGROUND INFORMATION: The parasitic protozoan Trichomonas vaginalis is the causative agent of trichomoniasis, a sexually transmitted disease. The phagocytic activity of this parasite has not been completely elucidated. In order to better understand the mechanisms of trichomonal phagocytosis, we have studied the in vitro capacity of T. vaginalis to phagocytose and degrade Saccharomyces cerevisiae cells. RESULTS AND CONCLUSIONS: To analyse the phagocytic ability and capacity, two isolates of T. vaginalis presenting different virulence grades were used. Complementary techniques, such as fluorescence microscopy, computer-based fluorescence analysis, scanning and transmission electron microscopy and the use of drugs that interfere with the actin microfilaments, were used in order to follow the behaviour of the actin cytoskeleton during phagocytosis of yeast cells by T. vaginalis. It was concluded that: (1) T. vaginalis changes its shape rapidly and engulfs the yeast cells, which are almost as large as the parasite; (2) long-term and fresh cultures are able to phagocytose, although the low-virulence strain JT demonstrated a lower activity when compared with the highly virulent T016 isolate; (3) the T016 strain exhibited an amoeboid morphology during the internalization of yeast cells in contrast with the JT strain; (4) attachment of yeast cells to the parasite occurs via the whole cell surface, including both anterior and recurrent flagella; (5) two forms of phagocytosis were observed: a 'sinking' process without any apparent participation of plasma membrane extensions and the classical phagocytosis where pseudopodia are extended toward the target cell; (6) the internalized S. cerevisiae are digested in lysosomes; (7) competitor sugars D-mannose or L-fucose inhibit the phagocytosis, and inhibition was 1.67 times higher in long-term cultured JT than that of the parasites from fresh isolate T016; (8) a thick layer of actin microfilaments was present underlying the plasma membrane, and especially in the pseudopodia and around the phagocytosed particles; (9) a dramatic change in the distribution pattern of fibrillar actin occurred during phagocytosis; (10) cytochalasin D depressed the phagocytosis; (11) a non-specific recognition and phagocytosis of yeast cells by T. vaginalis is mediated by a mannose receptor present on the parasite surface; (12) the phagocytic process may occur simultaneously during mitosis of the parasite.     

乳酸杆菌的代谢产物对体外培养的阴道毛滴虫的影响

目的研究乳酸杆菌代谢产物对体外培养的阴道毛滴虫的杀伤作用。方法检测不同浓度的乳酸杆菌代谢产物对体外培养的阴道毛滴虫在不同作用时间下的杀伤率。结果乳酸杆菌代谢产物浓度10%,作用时间2 h、4 h和6 h体外培养毛滴虫的杀伤率分别为26.43%、37.47%和46.35%;浓度25%时杀伤率分别为43.56%、74.65%和90.15%;浓度50%杀伤率分别为92.36%、95.23%和99.01%。结论乳酸杆菌代谢产物的浓度越高,对体外培养的毛滴虫的杀伤力越大,作用时间越长,效果越好。     

Excretory-Secretory Products of Trichomonas vaginalis Cause Apoptosis in Mouse Sperm in Vitro

Excretory-secretory products (ESP) of T. vaginalis have been shown to inhibit sperm motility, viability, and functional integrity, leading to a decreased fertilization rate in vitro. This study investigated whether T. vaginalis induce apoptosis and ultrastructural changes of sperm using flow cytometry and electron microscopy. Incubation of sperm with T. vaginalis ESP increased phosphatidylserine externalization and DNA fragmentation, and decreased mitochondrial membrane potential. Transmission electron microscopy of sperm incubated with ESP revealed abnormal features such as distorted heads, broken necks, and acrosomes exocytosis. This is the first report that demonstrates a direct impact of T. vaginalis ESP on sperm apoptosis and architecture in vitro.     

Proliferation of Mouse Prostate Cancer Cells Inflamed by Trichomonas vaginalis

Our objective was to investigate whether inflammatory microenvironment induced by Trichomonas vaginalis infection can stimulate proliferation of prostate cancer (PCa) cells in vitro and in vivo mouse experiments. The production of CXCL1 and CCL2 increased when cells of the mouse PCa cells (TRAMP-C2 cell line) were infected with live T. vaginalis. T. vaginalis-conditioned medium (TCM) prepared from co-culture of PCa cells and T. vaginalis increased PCa cells migration, proliferation and invasion. The cytokine receptors (CXCR2, CCR2, gp130) were expressed higher on the PCa cells treated with TCM. Pretreatment of PCa cells with antibodies to these cytokine receptors significantly reduced the proliferation, mobility and invasiveness of PCa cells, indicating that TCM has its effect through cytokine-cytokine receptor signaling. In C57BL/6 mice, the prostates injected with T. vaginalis mixed PCa cells were larger than those injected with PCa cells alone after 4 weeks. Expression of epithelial-mesenchymal transition markers and cyclin D1 in the prostate tissue injected with T. vaginalis mixed PCa cells increased than those of PCa cells alone. Collectively, it was suggested that inflammatory reactions by T. vaginalis-stimulated PCa cells increase the proliferation and invasion of PCa cells through cytokine-cytokine receptor signaling pathways.     

Chelex-100法及酚氯仿法提取阴道毛滴虫DNA的比较

目的-比较Chelex-100法和酚氯仿法提取阴道毛滴虫基因组DNA。方法-分别用Chelex-100法和酚氯仿法提取阴道毛滴虫基因组DNA,用PCR法检测DNA提取的有效性。结果两种方法提取的DNA经PCR扩增均有特定的条带。结论-两种方法均能提取阴道毛滴虫DNA。Chelex-100方法简便、省时,较适用于分子生物学研究及临床PCR扩增使用。