阴道毛滴虫在两种培养基中的生长与形态观察

目的通过对改良肝浸汤培养液及RPM11640培养基内阴道毛滴虫生长情况的观察，选择适合教学和科研的培养基。方法用临床分离的阴道毛滴虫标本，分别接种至改良肝浸液及RPM11640两种培养基内进行培养，pH值为5．6～5．8，通过计数了解生长情况并观察阴道毛滴虫的形态。结果肝浸汤培养液中培养48h，阴道毛滴虫达到生长高峰，虫体呈现出多分裂相，虫体变大、呈圆形，内含4～12个细胞核，细胞膜外缘可见数丛鞭毛。RPM11640培养基中阴道毛滴虫72h达到生长高峰，虫体密度约为肝浸液高峰期的2／3，未见多分裂现象，虫体与生理盐水涂片中阴道毛滴虫形态、大小相近。结论改良的肝浸液培养基适于阴道毛滴虫的药物试验及其他实验项目的研究，RPM11640培养基更适用于教学、标本的制作及实验室保种工作。     

3种不同培养基体外培养阴道毛滴虫效果的比较

目的　探索阴道毛滴虫体外培养的适宜条件。　方法　用临床分离的阴道毛滴虫 ,按 9.0× 10 4 / ml的接种量转种至 3种不同培养基进行培养 ,p H值为 5 .6。　结果　经 3种培养基培养 ,96 h后阴道毛滴虫数量存在差异 ,其中以半胱氨酸 -肝 -胨 -麦芽糖培养基 培养基 )虫数较多 ,肝 -胨 -麦芽糖培养基 培养基 )次之 ,大豆 -肝 -胨 -麦芽糖培养基 培养基 )较少。培养基 与培养基 及培养基 相比较 ,滴虫存活率差异具有显著性意义 P<0 .0 1,P<0 .0 5 ) ,滴虫生长密度差异也具有显著性意义 P<0 .0 1,P<0 .0 5 ) ,生长密度高峰持续时间分别为 192 h、14 4 h和 96h,最长存活时间分别为 2 88h、2 16 h和 192 h。　结论　半胱氨酸 -肝 -胨 -麦芽糖培养基较适于阴道毛滴虫体外增殖     

阴道毛滴虫在两种培养基中的生长与形态观察

目的 通过对改良肝浸汤培养液及RPMI1640培养基内阴道毛滴虫生长情况的观察,选择适合教学和科研的培养基。 方法 用临床分离的阴道毛滴虫标本,分别接种至改良肝浸液及RPMI1640两种培养基内进行培养,pH值为5.6~5.8,通过计数了解生长情况并观察阴道毛滴虫的形态。 结果 肝浸汤培养液中培养48 h,阴道毛滴虫达到生长高峰,虫体呈现出多分裂相,虫体变大、呈圆形,内含4~12个细胞核,细胞膜外缘可见数丛鞭毛。RPMI1640培养基中阴道毛滴虫72 h达到生长高峰,虫体密度约为肝浸液高峰期的2／3,未见多分裂现象,虫体与生理盐水涂片中阴道毛滴虫形态、大小相近。 结论 改良的肝浸液培养基适于阴道毛滴虫的药物试验及其他实验项目的研究,RPMI1640培养基更适用于教学、标本的制作及实验室保种工作。     

双氢青蒿素对阴道毛滴虫微丝作用的观察

目的　探讨双氢青蒿素对阴道毛滴虫的杀伤效果及作用机制。　方法　用含双氢青蒿素的肝浸汤培养基培养阴道毛滴虫 ,观察药物对滴虫的杀灭效果 ;用激光共聚焦显微镜观察双氢青蒿素作用前后滴虫微丝的变化。　结果　随药物作用时间延长和药物浓度增加 ,阴道毛滴虫死亡率增高。同一时间随着药物浓度的增高 ,虫体死亡率升高 (P <0 .0 1)。作用 6h ,双氢青蒿素 0 .6mg ml虫体死亡率是 2 0 % ,而 1.0mg ml时高达 85 % ;同一药物浓度随着作用时间延长 ,滴虫死亡率也升高 (P <0 .0 5 ) ,药物 0 .8mg ml时 ,6h、8h、10h、12h和 14h死亡率分别是 43 %、68%、86%、97%和 10 0 %。药物作用后滴虫微丝排列疏松 ,有空隙生成。排列杂乱无序。　结论　双氢青蒿素可作用于滴虫微丝结构 ,破坏微丝 ,具有较强的杀滴虫作用。     

在固体培养基中培养阴道毛滴虫的研究

目的 研究阴道毛滴虫在固态培养基中的生长条件。方法 厌养箱培养。结果 阴道毛滴虫在固态培养基中生长繁殖的最佳条件为0．45％琼脂浓度，37℃，厌氧箱培养。在直径6cm的培养皿中接种100个中虫数为宜。随着培养时间增加，虫落直径增大，至培养第5d直径最大，其后死亡率明显上升。阴道毛滴虫在固态培养基中最多可活8d。结论 阴道毛滴虫可在固态培养基中生存繁殖，此项技术为深入研究阴道毛滴虫提供了新的培养方法。     

低温保存阴道毛滴虫的实验观察

利用5%、10%、15%、20%、25%、30%浓度的二甲基亚砜和甘油肝浸汤培养液对临床分离的阴道毛滴虫进行4℃保存观察。二甲基亚砜和甘油两实验组最适保种浓度分别为15%和10%,50%虫体存活率天数分别为29和27 d,最长保存天数分别为31和29 d;而空白对照组仅为5和7 d。结果表明,15%二甲基亚砜或10%甘油肝浸汤培养液4℃冷藏保存阴道毛滴虫可显著延长其保存期。     

在固体培养基中培养阴道毛滴虫的研究

目的研究阴道毛滴虫在固态培养基中的生长条件.方法厌养箱培养.结果阴道毛滴虫在固态培养基中生长繁殖的最佳条件为0.45%琼脂浓度,37 C,厌氧箱培养.在直径6 cm的培养皿中接种100个虫数为宜.随着培养时间增加,虫落直径增大,至培养第5 d直径最大,其后死亡率明显上升.阴道毛滴虫在固态培养基中最多可活8 d.结论阴道毛滴虫可在固态培养基中生存繁殖,此项技术为深入研究阴道毛滴虫提供了新的培养方法.     

阴道毛滴虫经扁桃酰胺作用后的扫描电镜观察

扁桃酰胺是桃叶浸液中杀灭阴道毛滴虫的有效成分,作者对其杀虫机制进行过透射电镜观察,现将扫描电镜观察结果作一简报。 方法 自滴虫性阴道炎病人取材,接种于肝浸汤培养基中,37℃下稳定培养4代后实验。所用滴虫已培养48h,虫体生长旺盛、无死亡者进行实验。实验分两管,一为药物管(含0.8％扁桃酰胺);另一为     

MTT染色法计数阴道毛滴虫体外增殖密度的试验

目的评价四氮唑蓝(5-’diphenyl tetrazolium bromide,MTT)染色法计数阴道毛滴虫增殖密度的可行性。方法在已接种了阴道毛滴虫的肝浸汤(含小牛血清)、RPMI-1640(含小牛血清)、RPMI-1640(无血清)培养基中加入MTT,与血球计数板法进行比较。结果加入MTT后4、244、8 h,上述3种培养基中大部分阴道毛滴虫虫体内部未见蓝色结晶体形成,有血清的培养液镜下观察视野不清晰。结论MTT法不宜用于计数体外培养阴道毛滴虫的增殖密度。     

中药体外抗阴道毛滴虫的试验研究

为开辟新的抗阴道毛滴虫药物 ,本实验选择了 3种具有杀虫、消炎作用的中药进行了体外抗阴道毛滴虫的效果观察。1　材料和方法1.1　培养基　选用肝浸汤培养基 ,p H5 .5～ 6 .0。1.2　试验虫种　取初次确诊为滴虫性阴道炎患者阴道后穹窿处分泌物 ,接种于培养管内 ,每 3d转种传代 ,稳定培养三代开始试验。1.3　药物制备　取仙鹤草、苦参和雷丸各 10 g,置 2 0 0 ml蒸馏水中浸泡 6 h,煮沸 30 min取出药液 ,再加水 10 0 ml,微火煎沸2 0 min取出液体并与前者药液混合 ,8层纱布滤过 ,加热浓缩成2 0 ml,即为 5 0 %的药液。设对照组为培养液。1.4　药…     

组织贴块法人的支气管上皮细胞的原代培养

目的 通过组织贴块法建立人的支气管上皮细胞原代培养的方法.方法 采用无血清支气管上皮培养基,对经手术切除获得的支气管进行组织贴块法培养获得的细胞进行原代培养和传代,通过倒置显微镜以及细胞免疫化学观察和鉴定细胞.结果 此方法获得的细胞成活率高、纯度高,细胞呈扁平、多边形,象铺路的鹅卵石样分布.细胞角蛋白表达阳性.结论 组织贴块法是一种简便、有效的培养人的支气管上皮细胞的方法,无血清培养基可提供满意的生长条件,提高上皮细胞的纯度,为进一步研究不同的呼吸系统疾病提供了良好的模型.     

Sore throat: to culture or not to culture

The traditional use of the throat culture to guide treatment of potential streptococcal pharyngitis recently has been questioned because of the following facts: antibiotic therapy within the first 48 hours of illness reduces the severity and symptoms of streptococcal pharyngitis; more rapid treatment of streptococcal pharyngitis may be more effective in prevention of rheumatic fever; clinical and Gram stain criteria can identify individuals with a high probability of a positive throat culture; new latex agglutination tests can identify within minutes individuals with a high probability of a positive throat culture; the throat culture may be falsely negative in individuals with partially treated pharyngitis, an inadequately swabbed or plated specimen, or an improperly read culture plate; and incomplete patient followup increases the true societal cost of traditional therapy based on throat culture results. These issues suggest that a sufficiently accurate screening test for guiding antibiotic therapy at the initial visit may be more clinically beneficial and cost effective than recalling all positive throat culture patients for antibiotic therapy. Nonetheless, a throat culture remains important when a screening test of a patient with suspected streptococcal pharyngitis is negative. Indications for culturing pharyngitis for other potential pathogens also are discussed.     

Serum-free culture of human hemopoietic progenitors in attenuated culture media

To elucidate the precise mechanisms of molecular and cellular regulation of hemopoiesis, it is necessary to develop a chemically defined culture assay for purified hemopoietic progenitors. To approach this long-term goal, we attempted to develop a serum-free culture system for enriched human progenitors that permits expression of all hemopoietic lineages and stages of development. Preliminary studies indicated that alpha-medium was superior to Iscove's modified Dulbecco's medium (IMDM) and that culture under low (5%) oxygen condition was better than an ambient level of oxygen. We developed an attenuated (modified quarter-strength) alpha-medium and compared the colony-supporting ability of the three media by plating 1,000 bone marrow null cells per dish in the presence of a combination of recombinant human colony-stimulating factors (CSFs). The numbers of colonies supported in alpha-medium and attenuated alpha-medium were approximately 70% of those in serum-containing cultures. IMDM failed to support colony formation. While, in general, the colony sizes were smaller in the serum-free cultures than in the serum-containing cultures, a variety of types of single lineage and multilineage colonies were seen in serum-free culture. A linear relationship between cell number and colony formation was seen in 100-2,000 cells per dish. Serum-free cultures of enriched human progenitors should be an important tool for analysis of the mechanisms of recombinant CSFs.     

Purification and culture of mouse gallbladder epithelial cells in secondary culture using microexplant culture on collagen gel]

We established a new method of purification and culture of mouse gallbladder epithelial cells. In primary culture, mouse gallbladder tissue was cultured on collagen gel using microexplant culture method; the epithelial cells spread in a single sheet from the microexplant toward the periphery on the collagen gel. Tiny fragment of the collagen gel containing the border of epithelial cell layer was retransplanted onto the another collagen gel for secondary culture. This retransplantation allowed the proliferation of the epithelial cells without mesenchymal contamination, which spread on the collagen gel at the rate of 0.29 +/- 0.06 mm per day for more than two weeks. The cultured epithelial cells showed cuboid or columnar shape with focal mucus production, which is morphologically and functionally similar to the in vivo epithelial cells. This suggests that this in vitro culture method can be used to pathophysiological studies of biliary epithelial cells as well as for the method of purification of the epithelial cells.     

Effect of Campylobacter jejuni extracts and culture supernatants on cell culture

This study reports the effect of culture supernatants and extracts of Campylobacter jejuni isolated in Bangladesh from patients with gastroenteritis, asymptomatic carriers, poultry, and animals on tissue culture system. Isolates from patients produced morphologic changes on HeLa, Vero and Y1 adrenal cell lines. Incorporation of polymyxin B in the culture medium enhanced the effect on cell lines significantly and sonicated extracts of the organisms had an even more significant effect. Almost all sonicated extracts of patients' isolates had effect on HeLa cells. Animal isolates, using identical conditions, had much less effect on cell lines whereas isolates from asymptomatic carriers produced almost no effect. This toxicity of the patients' isolates might indicate some relation to pathogenic strains.     

An optimal culture condition maintains human hepatocyte phenotype after long-term culture.

BACKGROUND: Long-term culture of primary hepatocytes from various species is impeded by a decrease of cell viability and a loss of hepatocyte-specific function. The aim of the present study was to investigate whether our optimal culture condition (OC) can maintain the phenotype of primary hepatocytes in long-term culture. METHODS: Primary human hepatocytes were cultured in either hepatocyte maintenance medium (HM) or OC for 2-4 weeks. Expression of hepatocyte-specific genes was determined by real-time quantitative RT-PCR. RESULTS: The level of albumin mRNA in human hepatocytes cultured in OC was 11-fold more than in HM and gene expression levels of alpha1-antitrypsin and transferrin were at approximately 40 and 11% of freshly isolated primary human hepatocytes. Electron microscopy revealed that cells in OC displayed hepatocyte properties (e.g. polarity, junctional complexes, bile canaliculi and glycogen particles). Cytochrome P4501A1/2 activity of hepatocytes cultured in OC was 15- and 17-fold higher than in HM at 2 and 4 weeks of culture, and DNA synthesis was higher. CONCLUSIONS: Using our optimal culture condition, we were able to maintain the phenotype of primary human hepatocytes in long-term culture. They not only maintain better liver-specific function, but also retain higher proliferative potential.     

MGIT960与罗氏培养法在结核分枝杆菌培养及药敏试验中的比对分析

目的 比对全自动Bactec MGIT960（MGIT960）系统与改良罗氏（L-J）培养方法 2者在结核分枝杆菌培养、鉴定及药敏试验中的优越性。 方法 按国家疾病预防控制中心（CDC）-碧迪项目辽宁工作方案要求和国际标准WS288-2009《肺结核诊断标准》纳入临床病例。本实验共纳入病例435例,其中男性319例,女性116例,平均年龄52.3岁（10～86岁）。上述每份病例标本均分别进行痰液涂片抗酸染色（萋-尼法）镜检,L-J固体培养和MGIT960系统液体培养。培养阳性者分别进行一线药物异烟肼（INH）、利福平（RIF）、链霉素（STR）和乙胺丁醇（EMB）的L-J法药敏试验（比例法）和MGIT960药敏试验;分别计算出抗酸染色镜检与MGIT960和L-J培养的符合率; L-J培养和MGIT960培养阳性检出率;L-J和MGIT960两种培养方法药敏试验所需的培养时间及符合率等。 结果MGIT960培养法和L-J培养法与痰涂片镜检的阳性符合率分别为90.8%（187/206）和89.3%（184/206）;MGIT960与L-J结果符合率92.8%（219/236）;MGIT960和L-J平均培养时间分别为9.5 d和31.5 d;MGIT960和L-J一线药物INH、RIF、STR和EMB敏感性试验的符合率分别为92.7%、95.9%、91.8%和97.3%。 结论 全自动MGIT960系统培养方法的敏感度和特异度高于L-J培养法,报告时间平均缩短至9.5-d,该系统是目前最为理想的快速结核分枝杆菌培养、鉴定和药敏试验检测系统。