原生质体紫外诱变技术在核糖菌种选育中的应用

本文采用原生质体紫外诱变技术,经过硫酸二乙酯(DES)诱变后筛选出的NO3菌。采用溶菌酶浓度0.8mg/ml,酶解温度36℃,酶解时间120min,获得原生质体;采用10s作为紫外原生质体诱变的作用时间,选育出一株稳定的核糖生产菌TJ3,该菌具有莽草酸营养缺陷,耐高糖、以核糖为唯一碳源不生长等遗传标记。

D-核糖生产菌的原生质体诱变育种及其发酵条件的研究

以短小芽孢杆菌为出发菌株 ,经过硫酸二乙酯和原生质体紫外诱变 ,定向选育出一株稳定的核糖生产菌 TJ3,该菌株具有莽草酸营养缺陷、耐高糖、以核糖为唯一碳源不生长等遗传标记。在发酵条件试验中 ,采用均匀设计理论 ,利用微机对试验数据进行统计处理 ,确定出以葡萄糖为原料直接发酵生产核糖的优化配比。在最适培养条件下 ,摇瓶发酵平均产核糖 30 .12 mg/m

卡介菌核糖核酸和人血丙种球蛋白防治小儿支气管哮喘的药物经济学评价

目的 :评价卡介菌核糖核酸和人血丙种球蛋白两种免疫调节剂对于小儿支气管哮喘防治的药物经济学。方法 :对 75例支气管哮喘患儿随机分组 ,分别应用卡介菌核糖核酸和静脉用人血丙种球蛋白防治 ,进行成本 -效果分析。结果 :两种药物对于防治小儿支气管哮喘各有特点 ,均可起到较好的防治效果。结论

高频电离治疗仪联合局部注射卡介菌核糖核酸治疗尖锐湿疣疗效观察

目的:探讨高频电离治疗仪联合局部注射卡介菌核糖核酸治疗尖锐湿疣(CA)的方法和疗效。方法:应用高频电离治疗仪切除疣体以及局部注射卡介菌核糖核酸(BCG-PSN)等治疗手段,对156例CA患者采取随机分组治疗的临床试验。结果:156例患者随访3～6个月。治疗组痊愈51例,显效20例,无效3例,复发4例,有效率为91.03%;对照组治愈27例,显效14例,无效15例,复发22例,有效率为52.56%。结论:高频电离治疗仪联合局部注射BCG-PSN治疗尖锐湿疗效满意,值得推广。

经尿道汽化电切联合卡介菌核糖核酸膀胱内灌注治疗腺性膀胱炎分析

目的探讨经尿道汽化电切联合卡介菌核糖核酸膀胱内灌注治疗腺性膀胱炎的方法和疗效。方法应用抗感染、经尿道汽化电切以及卡介菌核糖核酸膀胱内灌注等治疗手段,对98例腺性膀胱炎患者进行随机分组治疗。结果 98例患者随访6～24个月,平均随访12个月,其中治疗组治愈31例,好转20例,复发4例,有效率为92.7%;对照组治愈7例,好转17例,复发19例,有效率为55.8%。结论经尿道汽化电切联合卡介菌核糖核酸膀胱内灌注治疗腺性膀胱炎疗效满意。

高频电离治疗仪联合局部注射卡介菌核糖核酸治疗尖锐湿疣疗效观察

目的探讨高频电离治疗仪联合局部注射卡介菌核糖核酸治疗尖锐湿疣的方法和疗效。方法应用高频电离治疗仪切除疣体以及局部注射卡介菌核糖核酸等治疗手段,对156例尖锐湿疣患者采取随机分组治疗的临床试验。结果 156例患者随访3～6个月,平均随访4.3个月,其中治疗组痊愈51例,显效20例,无效3例,复发4例,有效率为91.03%;对照组治愈27例,显效14例,无效15例,复发22例,有效率为52.56%。结论高频电离治疗仪联合局部注射卡介菌核糖核酸治疗尖锐湿疗效满意。

高频电离治疗仪联合局部注射卡介菌核糖核酸治疗尖锐湿疣疗效观察

目的探讨高频电离治疗仪联合局部注射卡介菌核糖核酸治疗尖锐湿疣的方法和疗效。方法应用高频电离治疗仪切除疣体以及局部注射卡介菌核糖核酸等治疗手段,对156例尖锐湿疣患者采取随机分组治疗的临床试验。结果 156例患者随访3～6个月,平均随访4.3个月,其中治疗组痊愈51例,显效20例,无效3例,复发4例,有效率为91.03%;对照组治愈27例,显效14例,无效15例,复发22例,有效率为52.56%。结论高频电离治疗仪联合局部注射卡介菌核糖核酸治疗尖锐湿疗效满意,值得推广。

卡介菌多糖核酸对复发性单纯疱疹患者外周血T淋巴细胞亚群的影响

应用卡介菌多糖核酸治疗３２例复发性单纯疱疹患者，对治疗前、后Ｔ淋巴细胞亚群进行了检测。结果显示治疗前ＯＫＴ８细胞比正常人增多（Ｐ＜０．０２），ＯＫＴ４／ＯＫＴ８比值下降（Ｐ＜０．００２），治疗结束后ＯＫＴ４细胞比治疗前增多（Ｐ＜０．０２），ＯＫＴ４／Ｔ８比值升高（Ｐ＜０．００５）。提示复发性单纯疱疹患者细胞免疫功能已受核抑制，卡介菌多糖核酸具有较显著的免疫恢复和调节作用，对皮损复发有较好的控制。

枯草芽孢杆菌转酮醇酶变异株C1-B941菌种特性的初步研究

C1-B941是一株枯草芽孢杆菌(Bacillus subtilis)的转酮醇酶(Transketolase,TKT)变异株,与亲株C1相比,其在菌体形态及代谢特性上发生了显著变化,如对戊糖不利用,对葡萄糖等利用变弱,碳源利用存在抑制,表现为芳香氨基酸合成途径缺陷,对数期细胞聚集成链,孢子形成能力下降,在发酵培养基中可以积累D-核糖,发酵液具有广泛的用途。

特异性抗菌i-RNA预防烫伤大鼠金葡菌感染的作用及机理研究

本研究利用烧伤动物模型,对特异性抗菌免疫核糖核酸（i-RNA）预防烫伤大鼠感染的作用及机理进行了探讨。结果发现,治疗组经i-RNA治疗后所测得痂下组织细菌定量、心血细菌培养和伤后10d内存活率均明显优于对照组;并发现治疗组腹腔巨噬细胞（MΦ）吞噬功能、脾淋巴细胞转化率及诱生IL-2能力也显著高于对照组。提示特异性抗菌i-RNA对烧伤感染有一定的预防作用,可增强烧伤机体的免疫功能,促使其受抑制的免疫功能及早得以恢复。

中西医结合治疗扁平疣35例临床疗效观察

目的:观察卡介菌核糖核酸注射液配合火针治疗扁平疣的临床疗效。方法:将65例扁平疣患者,按治疗方法不同分为对照组30例和治疗组35例。对照组给肌注卡介菌多糖核酸注射液及外用干扰素软膏治疗;治疗组在对照组的基础上实施火针治疗,对两组患者临床疗效及皮损消退情况等进行观察。结果:治疗组总有效率为91.43%,明显优于对照组的66.67%,差异具有统计学意义(P<0.05)。结论:卡介菌核糖核酸注射液配合火针治疗扁平疣的疗效较好,值得临床推广。

A novel salt-tolerant Micrococcus sp. DUT_AHX capable of degrading nitrobenzene

A novel strain of Micrococcus sp.DUT_AHX,which was isolated from the sludge of a nitrobenzene(NB)-manufacturing plant and could utilize NB as the sole carbon source,was identified on the basis of physiological and biochemical tests and 16S ribosomal DNA(rDNA)sequence analysis.It can grow at the temperature up to 40℃or in the presence of NaCl concentration up to 12 g/L in Luria-Bertani(LB)medium.The optimal degradation conditions are as follows:temperature 37℃,pH 7.0,and shaking speed 150 r/min.The strain involves a partial reductive pathway due to the release of ammonia and can also utilize 2-aminophenol as the sole carbon source.Furthermore,the enzyme activity tests show that crude extracts of NB-grown strain DUT_AHX mainly contain 2-aminophenol 1,6-dioxygenase activity.The exploitation of salt-tolerant bacteria will be a remarkable improvement in NB bioremediation and wastewater treatment at high salinity and high temperature.

Diarrhoea-predominant irritable bowel syndrome distinguishable by 16S rRNA gene phylotype quantification

AIM:To study whether selected bacterial 16S ribosomal RNA(rRNA)gene phylotypes are capable of disting- uishing irritable bowel syndrome(IBS). METHODS:The faecal microbiota of twenty volunteers with IBS,subdivided into eight diarrhoea-predominant (IBS-D),eight constipation-predominant(IBS-C)and four mixed symptom-subtype(IBS-M)IBS patients,and fifteen control subjects,were analysed at three time-points with a set of fourteen quantitative real-timepolymerase chain reaction assays.All assays targeted 16S rRNA gene phylotypes putatively associated with IBS,based on 16S rRNA gene library sequence analysis. The target phylotypes were affiliated with Actinobac-teria,Bacteroidetes and Firmicutes.Eight of the target phylotypes had less than 95%similarity to cultured bacterial species according to their 16S rRNA gene sequence.The data analyses were made with repeated-measures ANCOVA-type modelling of the data and principle component analysis(PCA)with linear mixed-effects models applied to the principal component scores. RESULTS:Bacterial phylotypes Clostridium cocleatum 88%,Clostridium thermosuccinogenes 85%,Coprobacillus catenaformis 91%,Ruminococcus bromii-like, Ruminococcus torques 91%,and R.torques 93%were detected from all samples analysed.A multivariate analysis of the relative quantities of all 14 bacterial 16S rRNA gene phylotypes suggested that the intestinal microbiota of the IBS-D patients differed from other sample groups.The PCA on the first principal component(PC1),explaining 30.36%of the observed variation in the IBS-D patient group,was significantly altered from all other sample groups(IBS-D vs control, P=0.01;IBS-D vs IBS-M,P=0.00;IBS-D vs IBS-C, P=0.05).Significant differences were also observed in the levels of distinct phylotypes using relative values in proportion to the total amount of bacteria.A phy- lotype with 85%similarity to C.thermosuccinogenes was quantified in significantly different quantities among the IBS-D and control subjects(-4.08±0.90 vs -3.33±1.16,P=0.04)and IBS-D and IBS-M subjects (-4.08±0.90 vs-3.08±1.38,P=0.05).Furthermore,a phylotype with 94%similarity to R.torques was more prevalent in IBS-D patients'intestinal micro- biota than in that of control subjects(-2.43±1.49 vs -4.02±1.63,P=0.01).A phylotype with 93%simi- larity to R.torques was associated with control sam- ples when compared with IBS-M(-2.41±0.53 vs -2.92±0.56,P=0.00).Additionally,a R.bromii-like phylotype was associated with IBS-C patients in com- parison to control subjects(-1.61±1.83 vs-3.69± 2.42,P=0.01).All of the above mentioned phylotype specific alterations were independent of the effect of time. CONCLUSION:Significant phylotype level alterationsin the intestinal microbiotas of IBS patients were observed,further emphasizing the possible contribution of the gastrointestinal microbiota in IBS.

Isolation and identification of a bacteria strain Enterobacter cloacae I7 for degradation of hydrolyzed polyacrylamide

A bacteria strain for the degradation of hydrolyzed polyacrylamide （HPAM） was isolated from a curing pot in HPAM distribution station of Daqing Oilfield using Hungate anaerobic technique. The isolate was investigated from morphological, physiological, biochemical and molecular characterization. It is a Gram-negative, shortbacillus, non-spore-forming anaerobic bacteria with an optimum growth at 8.0 pH at 40℃. It can reduce sulfate to I-I2S. Alignment of 16S ribosomal DNA and 16S-23S ribosomal DNA intergenic spacer sequences suggests that this isolate is closely related to the Enterobacter cloacae. The isolate is identified as a new strain belonging to Enterobacter genus, temporarily named as Enterobacter cloacae 17. Analysis results of infrared spectroscopy （IR） show that the bacteria can use HPAM as the only carbon source, change the structure of HPAM polymer surface, and realize the hydrolysis of amide to carboxyl group by hydrolysis mechanism. It can degrade the side chain and change some functional groups, which obviously decreases the viscosity. GC-MS analysis indicates that the determined low-molecular weight degradation products of HPAM are polyacrylamide fragments with duplet bond, epoxy as well as carbonyl group, but most of them are acrylamide oligomer derivatives.

迪苏联合蓝科肤宁治疗面部激素依赖性皮炎疗效观察

目的:观察迪苏联合蓝科肤宁治疗面部激素依赖性皮炎的疗效。方法:86例患者随机分成2组,治疗组与对照组均外用蓝科肤宁溶液湿敷,治疗组加用迪苏,共5周,分别观察两组的疗效。结果:治疗组和对照组的总有效率分别为87.5%和57.9%,x^2=3.124,P<0.01,治疗组疗效优于对照组。结论:迪苏联合蓝科肤宁治疗面部激素依赖性皮炎疗效满意。

Bacillus Licheniformis Dhs-40, a Flocculating Active Strain and Its Flocculating Activity

Microbial flocculants have become a hot spot in recent years. A bacterial strain with high flocculating activity was isolated from seawater samples. The strain was defined as Bacillus licheniformis dhs-40 by 16S rDNA identification and Biolog test. Ultrasonication test confirmed the flocculating activity of the strain was both in fermentation supernatant and cell. According to flocculating activity curve, the ideal fermentation time for collecting flocculating active substances was two days. The flocculating activity of the strain was sensitive to pH. The strain could only preserve flocculating activity while pH varied from 7 to 11. However, it could preserve flocculating activity while temperature varied from -20℃to 100 ℃ Saccharide, protein, lipid, nucleic acids qualitative test and RNase, Proteinase K treatment confirmed the flocculating active substances were proteins. Their flocculating activities were insensitive to Proteinase K.

Discovery of thiazostatin siderphores from Streptomyces sp. HMU0027 with a genomic DNA-based PCR assay targeting nonribosomal peptide synthetase

Nonribosomal peptides （NRPs） represent a large family of natural products with great diversities of chemical structures and biological activities. The peptide backbones of NRPs are synthesized by nonribosomal peptide synthetases （NRPSs） that minimally consist of one adenylation （A） domain, one peptidyl carrier protein （PCP） and one condensation （C） domain. In this study, we carded out a PCR screening and identified 21 ＂positive＂ strains from 100 actinomycete strains with the degenerate primers designed from the conserved sequences of A domains of NRPSs. One of the 21 ＂positive＂ strains, Streptomyces sp. HMU0027, was selected for large-scale fermentation （9 L） based on HPLC analysis, and subsequent isolation and structural elucidation afforded seven NRPS-synthesized thiazostatin siderophore analogues （1-7）. Compound 1 was a new compound containing an unusual linkage between a phenolate siderophore and a sugar moiety. These results laid the foundation for further biosynthetic research of these thiazostatin analogues and highlighted the power of genome mining technologies based on biosynthetic knowledge in NRP discovery.

Simultaneous extraction of DNA and RNA from Escherichia coli BL 21 based on silica-coated magnetic nanoparticles

The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many commercial kits and methods are currently available that allow the extraction of only one type of nucleic acids-DNA or RNA. However, in parallel clinical detection of several diseases, a method for simultaneous extraction of both DNA and RNA from the same source is needed in such cases. In this study, a method for simultaneous extraction of DNA and RNA from bacteria based on magnetic nanoparticles(MNPs) was described. Lysis buffers were prepared to help the nucleic acid released and adsorbed to MNPs. Then, two washing buffers were used to remove the contamination of proteins and carbohydrates. The nucleic acids were finally eluted by Deoxyribonuclease(DNase) and Ribonucleases(RNase) free water. Different factors which might affect the purification of the nucleic acid were investigated, and the quantity and quality parameters of the nucleic acid were also recorded. The DNA and RNA extracted from bacteria were then respectively subjected to polymerase chain reaction(PCR) and reverse transcription PCR(RT-PCR) to further confirm its quality. The results indicated that our method can be successfully used to simultaneously extract DNA and RNA from bacteria.