布拉氏酵母菌联合维生素D对产后抑郁症干预的研究

目的 探讨布拉氏酵母菌联合维生素D治疗产后抑郁症的临床效果。方法 选择我院收治的产后抑郁症患者102例,随机分为观察组和对照组各51例。两组患者均给予心理干预,对照组予维生素D 800 IU/次,1次/d口服,观察组在对照组基础上联合布拉氏酵母菌0.5 g/次,2次/d口服,4周一疗程。观察两组患者临床疗效及心里干预效果。结果 观察组患者治愈率及总有效率均高于对照组,两组比较差异有统计学意义（P<0.01,P<0.05）。治疗前两组患者HAMD评分及1,25（OH）2D3水平比较,差异无统计学意义（P＞0.05）。治疗后两组患者HAMD评分均较治疗前下降,1,25（OH）2D3水平比治疗前升高,但观察组降、升幅度明显大于对照组（P＜0.01）。结论 布拉氏酵母菌联合维生素D干预产后抑郁症具有明显的临床疗效。     

布拉酵母防治病原微生物感染的机制及效果

肠杆菌科细菌、霍乱弧菌、难辨梭菌、幽门螺杆菌和原虫等病原体感染是引起腹泻的主要原因.布拉酵母作为一种益生菌,通过抗微生物、免疫调节、肠道屏障、增加肠道营养等方面维持肠道微生态平衡.该酵母用于治疗肠杆菌科细菌、霍乱弧菌等病原菌感染引起的急性腹泻,能显著降低腹泻频率,缩短腹泻时间;而作为辅助药物联合抗生素治疗难辨梭菌、幽门螺杆菌和原虫等顽固性病原微生物感染,具有提高根除率、减少副作用、降低复发率的作用.本研究综述了布拉酵母对若干腹泻病原体的防治机制和临床应用研究进展.     

硝呋太尔制霉素阴道软胶囊联合布拉酵母对细菌性阴道病的疗效及安全性评价

目的 探讨硝呋太尔制霉素阴道软胶囊联合布拉酵母对细菌性阴道病（BV）的临床疗效及安全性。方法 将90例门诊收治的BV患者随机分为治疗组与对照组各45例,对照组患者采用硝呋太尔制霉素阴道软胶囊1粒（500 mg）,清洁外阴后放入阴道深部,每晚一次。观察组患者在对照组基础上联合布拉酵母0.5 g/次,2次/d,口服,2周为一个疗程。比较两组患者临床疗效、复发率及不良反应发生率。结果 观察组患者总有效率显著高于对照组（93.33％ vs 77.77％）,复发率显著低于对照组（60.00% vs 12.00）,差异均有统计学意义（P＜0.05）。两组患者不良反应发生率差异无统计学意义（17.78% vs 15.56%,P>0.05）。结论 阴道内用药联合口服益生菌对BV患者的疗效优于单一阴道内给药,患者复发率低,安全性较高。     

Storage stability and sourdough acidification kinetic of freeze-dried Lactobacillus curvatus N19 under optimized cryoprotectant formulation

In this study, the response surface methodology was used to optimize the cryoprotective agent (skimmed milk powder, lactose and sucrose) formulation for enhancing the viability of Lactobacillus curvatus N19 during freeze-drying and storage stability of cells freeze-dried by using optimum formulation was evaluated. Our results showed that the most significant cryoprotective agent influencing the viability of L. curvatus N19 to freezing and freeze-drying was sucrose and skim milk, respectively. The optimal formulation of cryoprotective agents was 20 g/100 mL skim milk, 3.57 g/100 mL lactose and 10 g/100 mL sucrose. Using the optimum formulation during freeze-drying, the cell survival was found more than 98%. Under the optimal conditions, although only storage of the cells at 4 °C for 6 month retained the maximum stability (8.85 log cfu/g), the employed protectant matrix showed promising results at 25 °C (7.89 log cfu/g). The storage stability of cells under optimized conditions was predicted by accelerated storage test, which was demonstrated that the inactivation rate constant of the freeze-dried L. curvatus N19 powder was 9.74 × 10⁻⁶ 1/d for 4 °C and 2.08 × 10⁻³ 1/d for 25 °C. The loss of specific acidification activity after the storage at 4 and 25 °C was determined.     

布拉酵母菌联合三联疗法根除幽门螺杆菌疗效研究

目的探讨布拉酵母菌（S．Boulardii）散剂联合标准三联疗法根除幽门螺杆菌（Helicobacterpylori,H．pylori）的有效性和安全性。方法将210例初次接受根除治疗的Hpylori阳性患者随机分为2组,A组：口服埃索美拉唑＋阿莫西林＋呋喃唑酮,每日2次,疗程10d;B组：与A相同的三联10d疗法,加s．Boulardii散剂口服500mg／次,2次／d,共用药14d。观察患者不良反应发生情况及对药物耐受情况,停药4周后检测H．pylori根除率。结果A、B组Hpylori根除率按方案（PP）分析分别为70．7％、86．3％,按意向性（ITT）分析分别为66．7％、83．8％,B组PP和ITT根除率均高于A组,差异有统计学意义（P〈0．05）;B组不良反应发生率低于A组（8．8％vs 28．3％,P〈0．05）;在药物耐受程度方面,B组明显优于A组（P〈0．05）。结论S．Boulardii散剂联合标准三联疗法作为初次根除Hpylori治疗时可提高根除率,降低不良反应,增加药物耐受程度。     

布拉酵母联合小剂量红霉素治疗小儿功能性消化不良的临床观察

目的 探讨布拉酵母联合小剂量红霉素治疗小儿功能性消化不良（FD）的临床疗效。方法 将92例FD患者随机分为对照组（n=46）和观察组（n=46）。对照组单纯用红霉素3~5 mg/（kg•d）,3次/d饭前服用。观察组在对照组治疗方法基础上联合布拉酵母0.25 g/次,2次/d,餐后口服,2周一疗程。随访6个月观察其复发率。结果 观察组总有效率（95.65%）明显高于对照组（76.09%）,差异有统计学意义（P<0.01）。2组患者治疗后HAMD评分均较治疗前下降,但观察组降低幅度显著大于对照组（P＜0.01）。观察组患者症状消失时间显著短于对照组,差异有统计学意义（P<0.01）。治疗后随访6个月,观察组患者复发率明显低于对照组（P<0.05）。结论 布拉酵母联合小剂量红霉素对小儿FD具有显著的临床疗效,患者的复发率较低。     

布拉酵母菌联合奥美拉唑阿莫西林克拉霉素治疗顽固性幽门螺杆菌感染的临床研究

目的 探讨布拉酵母菌联合奥美拉唑阿莫西林克拉霉素三联疗法对幽门螺杆菌(Helicobacter pylori,H.pylori)顽固性感染的治疗效果.方法 将120例H.pylori顽固性感染患者分成两组,分别采用奥美拉唑的三联疗法和布拉酵母菌联合奥美拉唑三联疗法治疗14 d.结果 两组患者治疗14d后,奥美拉唑三联组和布拉酵母菌联合奥美拉唑三联组的H.pylori清除率分别是94.6％和96.6％,两组间差异无统计学意义;在不良反应方面,奥美拉唑三联治疗组中发生16例,明显高于布拉酵母菌联合奥美拉唑三联组的5例(P＜0.05).结论 布拉酵母菌联合奥美拉唑三联治疗方案不仅具有良好的H.pylori清除效果,而且不良反应少,是治疗顽固性H.pylori感染患者比较好的方法.     

布拉氏酵母菌散剂对NASH大鼠的疗效及其作用机制探讨

目的观察口服布拉氏酵母菌散剂对NASH大鼠的疗效及其对sR—A的影响。方法42只雄性Sprague Dawley（SD）大鼠,随机分为正常对照组（NG,n=14）、模型组（MG,n=14）和干预组（TB,n=14）;正常组给予普通饲料喂养,模型组给予高脂饲料喂养,17周起干预组给予布拉氏酵母菌散剂灌胃,24周末将所有大鼠一并处死。比较各组血清内毒素、谷丙转氨酶、谷草转氨酶;计算非酒精性脂肪性肝病评分;采用免疫荧光法测定SR—A蛋白的表达,RealtimePCR法检测大鼠肝脏SR-A、TNF—αmRNA水平。结果与正常对照组相比,模型组的大鼠门冬氨酸氨基转氨酶（AST）、丙氨酸氨基转氨酶（ALT）均明显升高;干预组的大鼠AST、ALT与模型组相比均下降;模型组血清内毒素水平与正常对照组相比明显增加[（0．36±0．02）EU／mL vs（0．17±0．01）EU／mL,P〈0．05];而与模型组相比,干预组血清内毒素水平减少（0．22±0．01）EU／mL vs（0．36±0．02）EU／Ml,P〈0．05）。肝组织SR—A在正常对照组呈弥漫性表达,模型组肝脏的表达明显减少,干预组肝组织SR—A的表达较模型组升高。模型组的大鼠肝组织SR—AmRNA水平与正常组相比显著减少（0．52±0．32vs1．43±0．46,P〈0．05）;而与模型组相比,干预组的大鼠肝组织SR—AmRNA增加（0．87±0．34vs0．52±0．32。P〈0．05）。与正常组相比,模型组的大鼠肝组织TNF-αmRNA水平明显增加（1．56±0．35vs0．57±0．23,P〈0．05）;而与模型组相比,干预组的大鼠肝组织TNF—dmRNA水平减少（1．23±0．24vs1．564-0．35,P〈0．05）。结论布拉氏酵母菌散剂能够减轻肝脏脂肪变性及炎症程度,其机制可能与改善肠道菌群、减少肠源性内毒素、增加肝组织SR-A的表达有关。     

布拉酵母联合美沙拉嗪对溃疡性结肠炎患者氧化应激的影响及疗效观察

目的 探讨布拉酵母联合美沙拉嗪对溃疡性结肠炎（UC）患者氧化应激的影响及疗效。方法 选取2016年7月至2017年12月我院内科门诊治疗的活动期UC患者76例,随机分为观察组和对照组。对照组患者予以美沙拉嗪肠溶片1.0 g/次,4次/d,口服。观察组在对照组基础上加用布拉酵母0.5 g/次,2次/d,连用8周。观察并比较两组患者治疗前后的血清一氧化氮（NO）、氧化低密度脂蛋白（ox-LDL）、谷胱甘肽过氧化物酶（GSH-Px）和超氧化物歧化酶（SOD）水平的变化,并评估患者临床疗效和不良反应。结果 治疗8周后,两组患者血清NO和ox-LDL水平较治疗前明显下降,GSH-Px和SOD水平较前明显上升（P0.05）。结论 布拉酵母联合美沙拉嗪肠溶片治疗UC患者的效果确切,安全性较高,其作用机制可能与其能降低血清NO和ox-LDL水平,升高血清GSH-Px和SOD水平,减轻氧化应激对肠黏膜损伤密切相关。     

布拉酵母菌对胃黏膜上皮细胞损伤的保护作用及机制

目的 研究布拉酵母菌对胃黏膜上皮细胞损伤的保护作用。方法 建立人胃黏膜上皮细胞GES-1单层细胞模型,并将其分为对照组,布拉酵母菌上清培养组,阿司匹林处理组（18.32 mmol/L）,阿司匹林+布拉酵母菌上清液组。采用流式细胞术检测各组细胞凋亡率;采用免疫荧光、Western blot方法检测胃黏膜上皮细胞中Occludin和ZO-1蛋白、JNK蛋白、ERK蛋白、磷酸化JNK（p-JNK）蛋白、磷酸化ERK（p-ERK）蛋白表达水平。结果 流式细胞术显示阿司匹林+布拉酵母菌上清液组凋亡率低于阿司匹林处理组,差异有统计学意义（P<0.05）;免疫荧光结果显示,阿司匹林+布拉酵母菌上清液组Occludin蛋白、ZO-1蛋白表达较阿司匹林处理组增加,Western blot结果显示,阿司匹林+布拉酵母菌上清液组的该两种蛋白表达量较阿司匹林处理组增加,阿司匹林+布拉酵母菌上清液组p-ERK、p-JNK蛋白表达较阿司匹林处理组减少。结论 布拉酵母菌对阿司匹林造成的胃黏膜上皮细胞损伤有保护作用,其机制可能通过抑制MAPK信号通路中ERK、JNK的激活而上调Occludin和ZO-1蛋白的表达。     

布拉氏酵母菌联合阴道用药治疗复发性外阴阴道假丝酵母菌病的临床疗效及安全性

目的 探讨布拉氏酵母菌联合阴道用药治疗复发性外阴阴道假丝酵母菌病（RVVC）的临床疗效及安全性。方法 将98例RVVC患者随机分为观察组和对照组各49例,两组均给予硝酸咪康唑栓0.2 g/粒联合重组人干扰素α2b栓10万IU/粒,于月经干净3 d后阴道给药,每次1粒,1次/d。观察组联合布拉氏酵母菌0.5 g/次,2次/d,口服。连续用药3个月经周期,比较两组临床疗效、治疗前及治疗后3、6个月阴道分泌物炎性指标水平、复发情况及不良反应等。结果 观察组临床总有效率明显高于对照组（χ2=4.305,P0.05）。治疗后3个月、6个月两组IL-6水平逐渐下降,IFN-γ水平逐渐上升,但观察组升降幅度明显高于对照组（P<0.01）。结论 布拉氏酵母菌联合阴道用药治疗RVVC的临床疗效显著,能有效改善阴道炎症因子水平,安全有效且复发率低。     

布拉酵母菌散联合蒙脱石散对轮状病毒肠炎患儿细胞免疫功能的影响及疗效观察

目的探讨布拉酵母菌散联合蒙脱石散对轮状病毒肠炎患儿细胞免疫功能的影响及疗效。方法76例婴幼儿轮状病毒肠炎随机分为观察组和对照组。两组患儿均予以补液、抗病毒、纠正水电解质及酸碱平衡紊乱等常规治疗。观察组患儿加用布拉酵母菌散剂和蒙脱石散治疗,对照组患儿予以单纯的蒙脱石散治疗。观察两组患儿治疗后临床疗效及不良反应,并比较两组患者治疗后1个月T淋巴细胞亚群的变化。结果治疗72h后,观察组患儿的临床总有效率为94．74％,明显高于对照组的78．95％（χ2=4．15,P〈0．05）,两组患儿治疗中未发现明显药物不良反应。治疗后1个月,观察组患儿CD4＋及CD4＋／CD8＋比值较治疗前明显上升,CD8＋较治疗前明显下降（P〈0．05）。而对照组患儿治疗前后CD4＋、CD8＋、CD4＋／CD8＋比值无明显变化（P〉0．05）。结论布拉酵母菌散联合蒙脱石散治疗婴幼儿轮状病毒肠炎具有较好的疗效,安全性较好,与增强患儿的细胞免疫功能密切相关。     

布拉酵母菌散联合早期微量喂养对早产儿喂养不耐受及肠道菌群的影响

目的 探讨布拉酵母菌散联合早期微量喂养对早产儿喂养不耐受及肠道菌群的影响。方法 选取2014年1月至2018年6月在我院住院治疗的喂养不耐受早产儿80例,随机分为2组,各40例。对照组早产儿生后早期间歇持续微量喂养（12~24 h内）,观察组早产儿在对照组基础上加用布拉酵母菌散0.125 g/次,1次/d,通过奶瓶或鼻饲给药。两组患儿均连用3周。观察两组早产儿治疗后恢复情况、肠道菌群变化及治疗效果。结果 观察组早产儿呕吐腹胀消失时间、达全胃肠喂养时间及恢复出生体重时间均明显短于对照组（均P<0.05）。治疗3周后,两组早产儿肠道乳杆菌和双歧杆菌数量较治疗前明显上升（均P<0.05）,且观察组早产儿上升幅度较对照组更大（均P<0.05）。治疗后观察组早产儿临床总有效率明显高于对照组,差异有统计学意义（95.0% vs 80.0%,χ2=4.16,P<0.05）。结论 布拉酵母菌散联合早期微量喂养治疗早产儿喂养不耐受的疗效显著,能加快患儿的恢复,其作用机制可能与其能促进肠道正常菌群定植,加快肠道菌群的建立密切相关。     

布拉氏酵母菌联合加味逍遥散治疗小儿功能性再发性腹痛的临床疗及安全性

目的观察布拉氏酵母菌联合加味逍遥散治疗小儿功能性再发性腹痛(Recurrent abdominal pain,RAP)的临床疗效和安全性。方法将66例5~14岁RAP患儿随机分为治疗组和对照组各33例,2组腹痛发作时适当应用解痉剂及对症处理。治疗组用加味逍遥散水煎服,每天一剂,分3次口服;联合布拉氏酵母菌250 mg/次,每天2次口服。对照组单独用加味逍遥散(用法同上),2周为一个疗程。结果治疗组显效率与总有效率明显高于对照组(P0.05)。结论布拉氏酵母菌联合加味逍遥散治疗儿童RAP疗效确切,不良反应小,依从性好。     

Dynamic 13C-tracer study of storage carbohydrate pools in aerobic glucose-limited Saccharomyces cerevisiae confirms a rapid steady-state turnover and fast mobilization during a modest stepup in the glucose uptake rate

In this research, two dynamic ¹³C-labeling experiments confirmed turnover and rapid mobilization of stored glycogen and trehalose in an aerobic glucose-limited chemostat ( D =0.05 h⁻¹) culture of Saccharomyces cerevisiae . In one experiment, the continuous feed to an aerobic glucose-limited chemostat culture of S. cerevisiae was instantaneously switched from naturally labeled to fully ¹³C labeled while maintaining the same feed rate before and after the switch. The dynamic replacements of naturally labeled intracellular glycolytic intermediates and CO₂ (in the off-gas) with their ¹³C-labeled equivalents were measured. The data of this experiment suggest that the continuous turnover of glycogen and trehalose is substantial ( c . 1/3 of the glycolytic flux). The second experiment combined the medium switch with a shiftup in the glucose feeding rate (dilution rate shiftup from 0.05 to 0.10 h⁻¹). This experiment triggered a strong but transient mobilization of storage carbon, that was channelled into glycolysis, causing a significant disruption in the dynamic labeling profile of glycolytic intermediates. The off-gas measurements in the shiftup experiment confirmed a considerable transient influx of ¹²C-carbon into glycolysis after the combined medium switch and dilution rate shiftup. This study shows that for accurate in vivo kinetic interpretation of rapid pulse experiments, glycogen and trehalose metabolism must be taken into account.     

The taxonomic position of Saccharomyces boulardii as evaluated by sequence analysis of the D1/D2 domain of 26S rDNA, the ITS1-5.8S rDNA-ITS2 region and the mitochondrial cytochrome-c oxidase II gene

A taxonomic study was carried out on eight strains of Saccharomyces boulardii. Morphological and physiological characteristics were consistent with those of Saccharomyces cerevisiae. Sequences of the D1/D2 domain of the 26S rDNA were identical for all strains examined and had a similarity value of 100% compared to sequences of the type strain of S. cerevisiae (CBS 1171T) and strain S288c. For all S. boulardii isolates was found the exact same ITS1-5.8S rDNA-ITS2 sequence, which displayed a close resemblance with the sequences published for S288c (99.9%), CBS 1171(T) (99.3%) and other S. cerevisiae strains. Sequence analysis of the mitochondrial cytochrome-c oxidase II gene (COX2) also resulted in identical sequences for the S. boulardii isolates and comparisons with available nucleotide sequences revealed close relatedness to strains of S. cerevisiae including S288c (99.5%) and CBS 1171(T) (96.6%). The electrophoretic karyotypes of the S. boulardii strains appeared quite uniform and although very typical of S. cerevisiae, they formed a cluster separate from strains of this species. The results of the present study strongly indicate a close relatedness of S. boulardii to S. cerevisiae and thereby support the recognition of S. boulardii as a member of S. cerevisiae and not as a separate species.     

冻干高活力纳豆芽胞杆菌菌粉保护剂的筛选和优化

【目的】对冻干高活力纳豆芽胞杆菌菌粉保护剂进行筛选和优化研究,提高菌粉活菌存活率。【方法】采用单因素实验和正交实验设计,通过测定活菌存活率,筛选出最佳保护剂的配方;并研究采用优化后冻干保护剂制备的菌粉在20°C、4°C、25°C下的保存稳定性。【结果】纳豆芽胞杆菌的有效保护剂是:脱脂乳粉、甘露醇、L-抗坏血酸钠。最佳冷冻干燥保护剂配方是:脱脂乳粉10%+甘露醇4%+L-抗坏血酸钠1%,存活率达到91.63%。菌粉在20°C、4°C、25°C下保存12个月后,存活率分别为:88.79%、70.16%和10.52%,说明菌粉在20°C和4°C下保存稳定性较好,25°C下稳定性比较差。【结论】对纳豆芽胞杆菌冻干菌粉保护剂的优化,对纳豆芽胞杆菌的应用、活菌产品的质量稳定及新产品的研发均有一定的指导意义。     

Comparative study of stability of soluble and cell wall invertase from Saccharomyces cerevisiae

Yeast Saccharomyces cerevisiae is the most significant source of enzyme invertase. It is mainly used in the food industry as a soluble or immobilized enzyme. The greatest amount of invertase is located in the periplasmic space in yeast. In this work, it was isolated into two forms of enzyme from yeast S. cerevisiae cell, soluble and cell wall invertase (CWI). Both forms of enzyme showed same temperature optimum (60°C), similar pH optimum, and kinetic parameters. The significant difference between these biocatalysts was observed in their thermal stability, stability in urea and methanol solution. At 60°C, CWI had 1.7 times longer half-life than soluble enzyme, while at 70°C CWI showed 8.7 times longer half-life than soluble enzyme. After 2-hr of incubation in 8?M urea solution, soluble invertase and CWI retained 10 and 60% of its initial activity, respectively. During 22?hr of incubation of both enzymes in 30 and 40% methanol, soluble invertase was completely inactivated, while CWI changed its activity within the experimental error. Therefore, soluble invertase and CWI have not shown any substantial difference, but CWI showed better thermal stability and stability in some of the typical protein-denaturing agents.     

pH and thermal stability studies of carboxymethyl cellulase from intergeneric fusants of Trichoderma reesei/Saccharomyces cerevisiae

The combined effect of pH and temperature on carboxymethyl cellulase from two intergeneric fusants (M 14 and M 62) of Trichoderma reesei QM 9414/Saccharomyces cerevisiae NCIM 3288 was studied using response surface methodology. A central composite design for two variables was employed for the optimization studies. This study was compared with similar studies carried out with Trichoderma reesei QM 9414. The optimal pH and temperature for the enzymes derived from these organisms were: for the fusant M 14—pH 5.7 and 41.7°C, for the fusant M 62—pH 5.3 and 43°C, and for Trichoderma reesei QM 9414—pH 4.31 and 38.3°C. Received 5 May 1997/ Accepted in revised form 17 July 1998     

Cloning and optimization of induction conditions for mature PsaA (pneumococcal surface adhesin A) expression in Escherichia coli and recombinant protein stability during long-term storage

The gene corresponding to mature PsaA from Streptococcus pneumoniae serotype 14 was cloned into a plasmid with kanamycin resistance and without a purification tag in Escherichia coli to express high levels of the recombinant protein for large-scale production as a potential vaccine candidate or as a carrier for polysaccharide conjugation at Bio-Manguinhos/Fiocruz. The evaluation of induction conditions (IPTG concentration, temperature and time) in E. coli was accomplished by experimental design techniques to enhance the expression level of mature recombinant PsaA (rPsaA). The optimization of induction process conditions led us to perform the recombinant protein induction at 25°C for 16 h, with 0.1mM IPTG in Terrific Broth medium. At these conditions, the level of mature rPsaA expression obtained in E. coli BL21 (DE3) Star by pET28a induction with IPTG was in the range of 0.8 g/L of culture medium, with a 10-fold lower concentration of inducer than usually employed, which contributes to a less expensive process. Mature rPsaA expressed in E. coli BL21 (DE3) Star accounted for approximately 30-35% of the total protein. rPsaA purification by ion exchange allowed the production of high-purity recombinant protein without fusion tags. The results presented in this work confirm that the purified recombinant protein maintains its stability and integrity for long periods of time in various storage conditions (temperatures of 4 or -70°C using different cryoprotectors) and for at least 3 years at 4 or -70°C in PBS. The conformation of the stored protein was confirmed using circular dichroism. Mature rPsaA antigenicity was proven by anti-rPsaA mouse serum recognition through western blot analysis, and no protein degradation was detected after long periods of storage.