The characterization of hCG regulation in cultured human amniotic fluid cells

The media from primary cultures and subcultures of second trimester human amniotic fluid (AF) cells were assayed by radioimmunoassay to quantitate production of human chorionic gonadotropin (hCG). Primary AF cultures produce more hCG per cell than do the corresponding subcultures. Sodium butyrate (2 mM) stimulates AF subcultures to produce 5-13 times more hCG per cell or per mg of cellular protein than do untreated subcultures. This stimulatory effect of sodium butyrate is dose dependent between 0 and 5 mM. Addition of sodium butyrate 24 hours after subculture, while stimulating production of hCG during the subsequent 3 days, also results in fewer cells and less protein per culture. This effect on cell growth is also dose-dependent. Previous investigators have proposed that the stimulation of hCG by sodium butyrate in other types of cell cultures is due to an effect of that agent on culture growth. Therefore, in these studies AF cells are allowed to grow to confluency before sodium butyrate was added. Production of hCG was stimulated by sodium butyrate about four-fold during the next 5 days although no significant changes were observed either in number of cells or amount of cellular protein per culture. These results suggest that stimulation of hCG by sodium butyrate is not dependent on its effect on growth of the cultures.     

影响微生物絮凝剂产生的因素研究

本文通过改变培养基的种类、培养基的碳源、氮源、无机盐离子来选择絮凝剂产生菌“Dfjm-1”高效低廉的培养基 ;通过改变培养基初始 p H值、培养温度、培养过程中的通气量等因素 ,得出“Dfjm-1”菌株产生絮凝剂的最佳条件。Dfjm-1产生絮凝剂的最佳因素为 :培养基初始 p H值为 6.5～ 7.0 ,培养温度为 3 0℃ ,培养时间为 60小时左右 ,通气量为 :早期 :2 50 r/ min;中期 1 50 r/ min;后期 1 0 0～ 1 50 r/ min     

木质素过氧化物酶和锰过氧化物酶对染料脱色的性能比较

通过控制培养基中Mn2+的浓度获得只含有木质素过氧化物酶(LiP)或锰过氧化物酶(MnP)的单一酶培养物,用于比较LiP和MnP对染料降解脱色的条件和能力的差异.结果表明,LiP对橙Ⅰ降解脱色的最佳条件为h2O₂0.15 mmol/L,pH 3.0,温度40 ℃;MnP为h2O₂0.15 mmol/L,pH 3.5,温度40 ℃.最佳条件下,考察LiP和MnP作用下染料浓度和染料类型对脱色的影响,发现LiP作用下脱色率明显高于MnP作用下的脱色率,表明LiP可作用的染料浓度比MnP高,可作用的染料范围比MnP大.由此认为LiP具有比MnP更强的脱色能力.      

海洋费氏弧菌培养条件的研究

以本室保存的一株海洋费氏弧菌作为实验对象,采用单因素试验方法研究培养基起始pH值、盐浓度和摇瓶装液量等因素对菌株生长的影响,利用分光光度法测定其生长曲线,采用均匀设计方法,对该菌的培养基组分进行优化研究,确定其适合的生长条件,结果表明:当培养温度为30℃,通气量为180r/min时,该菌株在20h左右达到生长稳定期,对甘油的利用率要优于葡萄糖,盐浓度在5%以下时生长良好,最适装液量为60mL/250mL,培养液初始pH值为8.0。确定100mL培养基配方为:蛋白胨2g,酵母粉1.0g,氯化铵1.2g,氯化钠0.2g,磷酸二氢钾1.0g,此条件下可以得到最大的生物量。     

Influence of glucose feeding on the ligninolytic enzyme production of the white-rot fungus Phanerochaete chrysosporium

The present work studied the influence of glucose feeding on the ligninolytic enzyme production of Phanerochaete chrysosporium in a nitrogen-limited (C/N ratio is 56/8.8 mmol/L) medium. Several sets of shaking flask experiments were conducted. The results showed that 2 g/L glucose feeding on the first day of the culture (24 h after the inoculation) simulated both fungal biomass growth and enzyme production. The manganese peroxidase (MnP) activity was 2.5 times greater than that produced in cultures without glucose feeding. Furthermore, the glucose feeding mode in fed-batch culture was also investigated. Compared to cultures with glucose feeding every 48 h, cultures with glucose feeding of 1.5 g/L (final concentration) every 24 h produced more enzymes. The peak and total yield of MnP activity were 2.7 and 3 times greater compared to the contrast culture, respectively, and the enzyme was kept stable for 4 days with an activity of over 200 U/L. Translated from Acta Scientise Circumstantiae, 2007, 27(3): 363–368 [译自: 环境科学学报]     

正十六烷微生物降解酶的定域和酶促降解性

以正十六烷为研究对象,通过室内实验,利用细胞静息技术提取了2株菌种的胞外酶、膜周酶及膜内酶对石油烃类污染物质的微生物降解酶定域,并研究了菌株受环境影响的产酶条件和酶的一般性质. 结果发现:蜡状芽孢杆菌DQ01能降解正十六烷的关键酶位于膜周和膜内,芽孢杆菌DQ02降解正十六烷的关键酶是胞外酶和膜内酶. 通过GC-MS对代谢产物进行测定发现,关键酶对十六烷的代谢途径是常见的单末端氧化. 2株菌种产酶的最佳环境条件:ρ(正十六烷)为100 mg/L,c(鼠李糖脂)为2 mmol/L. 另外,关键酶在pH为6.5～8.0的环境中活性较高,在pH为7.0左右时的活性最高. 酶促降解性的最适温度为30 ℃.      

Reduction of sera requirements in amniotic fluid cell culture

Reduction in serum requirement for culture of primary human amniotic fluid cells can be achieved by the addition of 10 growth-promoting factors to the nutrient medium. This supplemented medium preserves cell types normally found in amniotic fluid cell cultures supplemented with 20–30 per cent fetal bovine serum. The volume of amniotic fluid required to initiate culture can be as little as 1 ml. Amniotic fluid samples contaminated with red blood cells with no visible clot also grow well in the low serum medium. Cell-free amniotic fluid combined with equal parts of supplemented medium is useful in initiating cell culture.     

白腐真菌木质素降解酶的反应器发酵

为了获得更高的白腐真菌木质素降解酶的产量和相应的控制策略,应用5L搅拌罐生物反应器对氮限制下(C/N=56/2.2)P.chrysosporium产木质素降解酶进行了放大研究.结果表明,在分批发酵试验中,锰过氧化物酶(MnP)和漆酶(Lac)分别在培养第6d和第7d达到峰值,其酶活随时间的变化规律与摇瓶试验基本相同;而采用氮限制液体培养基进行补料没有获得更高的酶活,因此,可以得出采用发酵液体培养基作为补料液不利于白腐真菌持续产酶.另外,在分批发酵和分批补料发酵过程中,均发现体系pH值变化与白腐真菌生长和次生代谢产酶具有较好的相关性,当白腐真菌进入次生代谢阶段开始产酶时,体系pH值开始下降,随着发酵后期酶活的降低,pH值下降的幅度也逐渐变小,当体系酶活接近为0.0U/L时,pH值基本不再变化.因此,在实际发酵过程中,可以依据体系pH的变化间接了解白腐真菌的生长和产酶情况,而分批补料策略还有待于进一步研究.     

葡萄糖补料对白腐真菌P.chrysosporium产木质素降解酶的影响

为了提高木质素降解酶的产量,采用氮限制培养基(C/N=56.0/8.8),研究了葡萄糖补料对P.chrysosporium产木质素降解酶的影响.结果表明,在培养第1d即接种24h后进行葡萄糖补料,可以刺激菌丝体的生长和产酶,补料浓度达到2g·L-1时,补料体系中的MnP酶活可提高至空白对照样(不进行补料)的2.5倍.同时,对葡萄糖分批补料方式进行的试验结果显示,培养过程中每24h进行1次补料,葡萄糖补料终浓度为1.5g·L-1的补料方式,与每48h的补料方式相比,产酶效果更好.MnP酶活峰值和达到峰值时酶的总产量可提高至空白对照样(不进行补料)的2.7倍和3.0倍,且酶活能够在200U·L-1以上稳定4d.实验结果说明,葡萄糖补料可以有效刺激白腐真菌产酶,且连续低浓度葡萄糖补料可获得较优的产酶效果.     

Comparison of Multiple Passage Integrated Cell Culture-PCR and Cytopathogenic Effects in Cell Culture for the Assessment of Poliovirus Survival in Water

The goal of this study was to determine if a cytopathogenic effects (CPE) cell culture assay and an integrated cell culture PCR (ICC-PCR) assay would yield similar or different results when used to assess virus survival in water. Poliovirus type 1 was added to dechlorinated tapwater and stored at room temperature (22.5–24°C) for a total of 50 days. Samples were assayed at defined time intervals by the most probable number (MPN) method on Buffalo green monkey kidney cells (BGM) by CPE and additionally by ICC-PCR. Monolayers that were CPE negative on first passage were passed onto fresh monolayers of cells for a second and third time if still negative. By CPE assay, second passage was observed to yield a greater titer (2,300 vs. 24,000 MPN/ml) and third passage also resulted in an increased titer. ICC-PCR proved to be a more rapid and sensitive method than conventional cell culture for determining virus inactivation rates in water. Poliovirus survived in tapwater for up to 32 days, as assessed by both third passage ICC-PCR and CPE. There was no statistical difference in the inactivation rates between the two methods. To determine the total number of infectious viruses, these findings indicate the need for performing three cell culture passages or, alternatively, ICC-PCR on first passage.     

嗜麦芽窄食单胞菌J12对芘的降解特性

从前期实验中得到1株对芘具有降解能力的嗜麦芽窄食单胞菌,将其命名为J12。采用摇床振荡培养的方法,研究了不同实验条件(降解体系芘初始浓度、初始pH、投菌量)对J12降解芘的影响。结果表明:最佳降解条件为芘初始浓度20mg/L、投菌量2g/L、pH为7.0左右。在最佳降解条件下,研究了金属离子Fe3+、Mn2+、Mg2+对J12降解芘的影响,结果表明一定浓度Fe3+、Mn2+的投加对J12降解芘具有促进作用,而Mg2+则表现为轻微的抑制作用。向反应体系中添加鼠李糖脂,结果表明鼠李糖脂的添加对J12降解芘有一定的促进作用,其中在反应的后期促进作用较明显。对J12胞外酶和胞内酶粗酶液进行考察,结果表明在J12降解芘过程中起主要作用的为胞内酶,J12对芘的降解酶主要分布在胞内。     

Screening of flocculant-producing microorganisms and flocculating activity

A strain saccharomycete STSM-I with high flocculanting activity was isolated from activated sludge with conventional methods.The high production rate and the low cost STSM-I medium was obtained by selecting different kinds of media, carbon source, nitrogen source and inorganic salt ion. The best flocculant-producing conditions were found by changing medium initial pH, culture temperature and ventilation flow. The best flocculating effect was obtained by changing positive ion types, density and concentration of flocculant.     

一株芽胞杆菌和一株黄杆菌代谢芘的摄取方式解析

通过溶剂萃取、薄层色谱和紫外分光光度法研究了一株芽胞杆菌CN2和一株黄杆菌CN4在初始pH=6.00的条件下对芘的降解过程.研究表明:芘降解液的pH值随降解时间向中性波动变化;用牛肉膏蛋白胨培养出的CN2菌体表面具有亲水性,CN4菌体表面具有疏水性,CN2表面疏水性在芘的降解过程中随时间有变大的趋势,CN4表面疏水性在芘的降解过程中随时间有变小的趋势;从芘降解的整个过程看,大部分时间CN2的降解比CN4的快,CN4菌体捕获的芘量比CN2要大,尤其在接种刚开始的一段时期较为明显;细胞表面、细胞壁中、细胞内部均有芘富集迹象,其中细胞表面富集的芘最多.根据试验结果推测,在菌株对芘的降解过程中吸附和降解同时发生;菌体对芘首先通过表面吸附或其它方式结合,随后溶人细胞壁中,跨过细胞膜进入细胞内被降解;这个传质过程很可能不是连续的,而是由细菌控制的主动的间歇的动态过程.     

The effect of oxygen tension on colony formation and cell proliferation of amniotic fluid cellsin vitro

The effects of reduced oxygen concentrations in the gas phase over the culture medium on colony formation and cell proliferation were investigated in high and low cell density primary and secondary cultures of amniotic fluid cells. Using two standard culture methods (25 cm² plastic flasks and Leighton type tubes) a significantly reduced culture time was observed at high cell density for mass cultures by incubation within a low oxygen tension gas phase (2.5 per cent to 7.5 per cent O₂) instead of conventional air (18 per cent O₂). At low cell density colony formation was significantly enhanced in cultures grown at reduced oxygen tension. Using gas permeable membranes as support, lowering the oxygen tension from 7.5 per cent to 2.5 per cent yielded an increase in plating efficiency of cells from approximately 5 per cent to 25 per cent, whereas plating efficiency was less than 2 per cent for cells grown at ambient 18 per cent O₂. It is suggested that low oxygen tension in the gas phase is an effective means of enhancing clonal growth in amniotic fluid cell cultures, thereby reducing both culture time and risk of culture failure.     

有效微生物群(EM)抑藻效应研究

通过细胞液体培养系统地考察了有效微生物群(EM)对3种常见"水华"藻类生长的抑制作用及适宜条件。实验结果表明:EM能显著抑制衣藻、四尾栅裂藻和盘星藻的生长,培养7d时,3种藻类生物量平均降低率分别达46 80%,99 07%和97 17%;培养温度、培养时间、EM投加量及培养基pH等对EM的抑藻作用均有不同程度的影响。EM发挥良好抑藻作用(绿藻)的适宜条件:培养温度为30℃,培养时间≤5d,EM投加量(V(EM)∶V(源水))为1∶1000～1∶10000,培养基pH为6 0～8 0。      

新型专性厌氧菌利用葡萄糖进行发酵产氢的实验研究

采用批式发酵法对厌氧产氢菌株Bacteria.P利用葡萄糖发酵,在底物浓度、初始pH值、接种比例等不同培养条件下的产氢能力进行了研究。结果表明:专性厌氧菌P是一种高效产氢的菌株,在葡萄糖质量浓度为10 g/L、初始pH为6.0、接种比例为1∶20时,发酵气体总产量和细胞干重达到最大值,分别为485 mL和0.836 g/L。     

杂食性鱼类排泄物中藻类光能活性研究

借助叶绿素荧光技术,通过对杂食性鱼类鲫鱼(Crucian Carp)和金鲫鱼(Gold Crucian Carp)摄食微囊藻(Microcysis aeruginosa)后排泄物的培养,从鱼类排泄物藻类叶绿素荧光活性的角度探讨了杂食性鱼类在控藻上的应用.结果表明,鱼类摄食对微囊藻生长及叶绿素荧光参数均有显著影响(P<0.05).鲫鱼组的叶绿素荧光参数ΦPSII从第3d开始,Fv/Fo、Fv/Fm、ETR和qP从第5d开始随培养时间的延长而增加,而NPQ始终呈下降趋势.金鲫鱼组的叶绿素荧光参数(Fv/Fo、Fv/Fm、ΦPSII、ETR、qP)在培养期间一直呈降低趋势.鲫鱼组的Chl a浓度与细胞密度在培养期间先下降再上升,最后恢复至对照组水平,且Chl a浓度同部分叶绿素荧光参数(Fv/Fo、Fv/Fm、ΦPSII)呈现极显著正相关(P<0.01);金鲫鱼组的Chl a浓度与细胞密度于实验期间均下降,实验期间一直低于对照组,且两参数均与叶绿素荧光参数(Fv/Fo、Fv/Fm、ΦPSII、ETR、qP)呈极显著正相关(P<0.01).可见,微囊藻经鲫鱼摄食后,其叶绿素光合及生长活性经过短暂的下降后会逐渐恢复,而金鲫鱼可有效降低微囊藻活性,但金鲫鱼作为一种观赏鱼类,不适宜在大面积水体放养.     

地层介质对垃圾渗滤液的pH缓冲性能研究

通过一维土柱穿透实验和静态实验,分别从空间和时间2方面研究了不同天然地层介质对垃圾渗滤液的PH缓冲性能.结果表明,细砂、粉砂和粘土的本底pH缓冲容量分别为79.9·pH-1 mmol/kg、207.5·pH-1 mmol/kg、456.4·PH-1 mmol/kg.受垃圾渗滤液污染后,除细砂外,随着时间的延长、离污染源距离的减小,每种介质总的pH缓冲容量都逐渐减小;其中碳酸钙缓冲体系的缓冲容量均逐渐增大;硅酸盐、阳离子交换缓冲体系的缓冲容量无明显变化;铝、氧化铁缓冲体系的缓冲容量逐渐减少,但土壤铝缓冲体系的缓冲容量则呈现出先增大后减少的趋势.细砂各个缓冲体系的缓冲容量随时、空变化均呈现出略微增大的趋势.4种介质对渗滤液的pH缓冲能力的顺序是:细砂<粉砂<粘土<土壤.土壤、粘土对垃圾渗滤液有较好的pH缓冲性能,同时对渗滤液中各种污染物的迁移,扩散也有较好的控制作用.     

紫外诱变法提高好氧反硝化菌降解性能的研究

采用紫外诱变方法对从污水处理厂活性污泥中筛得的1株好氧反硝化菌(YS)进行处理,得到1株突变菌株(TB),考察了诱变前后2菌株的理化性能及反硝化能力的变化.结果表明,菌株TB的反硝化能力得到改善.在相同实验条件下,菌株TB对硝酸根的去除能力可从菌株YS的87%提高至93%,培养基中的亚硝酸根浓度从212.48 mg.L-1减少到37.62 mg.L-1,去除率从15%快速提高至85%,表明菌株TB对亚硝酸根的去除能力大幅提高,且传代前后菌株TB对硝酸根和亚硝酸根的去除率变化不大,遗传稳定性较好.对两菌株的反硝化性能影响因素的研究比较发现菌株YS的最适条件为:接种量1.5%左右,pH为6,C/N比为10,碳源为葡萄糖;而菌株TB的最适条件为:接种量1.5%左右,pH为9,C/N比为10,碳源为琥珀酸钠.     

中肋骨条藻与锥状斯氏藻藻际细菌溶藻效应研究

通过向对数生长期的中肋骨条藻和锥状斯氏藻培养体系中分别加入体积分数1%和10%的2216E培养基,以未加入培养基的对照组和10%无菌组作为对照,研究藻际细菌的溶藻效应.结果表明,对照组、1%组及10%无菌组的中肋骨条藻和锥状斯氏藻生长未受抑制,而10%组的中肋骨条藻和锥状斯氏藻生长则受到显著的抑制(p0.001),在第96h时,藻细胞几乎都被裂解,藻细胞数、叶绿素a含量及叶绿素荧光效率(Fv/Fm)显著低于对照组、1%组及10%无菌组(p0.001),表明藻际细菌具有抑制微藻生长的作用.在整个溶藻过程中,中肋骨条藻10%组细菌丰度增加了约250倍,锥状斯氏藻10%组藻际细菌丰度增加了约300倍,藻际细菌丰度的剧增可能是溶藻的主要原因.作为溶藻效果的评价指标,叶绿素荧光效率要优于细胞形态的改变、藻细胞数量的变化及叶绿素a含量,是较理想的溶藻检验指标.