体外培养人胆囊上皮细胞的形态学及培养条件的优化

目的　探索人胆囊上皮细胞的最佳分离方法和适宜的体外生长条件 ,为深入研究胆囊的生理功能和相关疾病的病理机理奠定基础。方法　用Ⅳ型胶原酶消化及钝性刮离法分离胆囊上皮细胞 ,两步贴壁法纯化。比较层粘连蛋白、多聚赖氨酸、纤维连接蛋白等不同培养基质对传代细胞生长的影响。光镜及电镜下观察细胞的形态和超微结构。结果　每个胆囊可分离得到 ( 1～ 5)× 10 7个胆囊上皮细胞 ,细胞活性可达 90 %。细胞呈典型的扁平多角状、柱状形态 ,片状贴壁生长 ,上皮细胞特异性抗原CK19表达阳性。结论　Ⅳ型胶原酶消化法加钝性刮剥分离法和两步贴壁法可成功分离高活性、高纯度的人胆囊上皮细胞 ;纤维连接蛋白包被的培养器皿和含 2 0 %小牛血清及 10ng/mlhEGF的DMEM培养基有利于胆囊上皮细胞的体外培养     

益生菌干预骨质疏松症相关机制的研究进展

骨质疏松症是一种以骨量减低和骨组织微结构退化为特征的骨骼疾病。随着社会人口老龄化的进展,骨质疏松症的发病率将会越来越高,尤其是绝经期女性,其发生骨质疏松症的风险较男性更高。骨质疏松症所带来的问题日趋严重,其给社会及家庭带来巨大的负担,尤其是骨质疏松症导致的身体重要部位的骨折。目前骨质疏松症的治疗方法有限,药物的治疗虽然有一定成效,但是药物并不能彻底根治骨质疏松症且药物需长期使用,因此药物的副作用的问题不容忽视。在人类的肠道定植着数量繁多且对机体健康有着重要作用的菌群,其中益生菌的健康效应也逐渐被挖掘。目前研究发现益生菌对机体诸多的疾病有显著的疗效,其中益生菌和骨质疏松症也有着密切的关系。益生菌可以通过复杂的方式干预骨质疏松症,但其具体机制仍未阐明。本文通过相关的报道进行综述,并从(1)炎症因子;(2)基因、配体及信号通路;(3)破骨细胞;(4)钙离子吸收等方面探讨益生菌干预骨质疏松症的可能机制,为骨质疏松症的防治提供新的思路。     

肌肉组织工程的基础研究-不同培养条件对卫星细胞增殖的影响

研究不同培养条件对卫星细胞体外增殖的影响,寻找理想的培养条件。方法：通过MTT法绘制生长曲线,比较不同胎牛血清、不同血清浓度、不同培养液配方对卫星细胞增殖的影响。通过贴壁试验比较胶原和多聚赖氨酸对卫星细胞贴壁的影响。结果：不同的血清、不同的浓度以及不同的培养液配方对卫星细胞增殖影响明显。含有10％四季青胎牛血清的DMEM／F12(2:1)的培养液可以获得最高的增殖效率。胶原更有利于细胞的贴壁。结论：以本试验得出的培养条件可以使卫星细胞培养周期缩短1倍以上,为肌肉组织工程的研究创造了良好的条件。．     

益生菌联合三联疗法根除幽门螺杆菌的临床研究

幽门螺杆菌(helicobacter pylori,hp)从发现至今已有20多年的历史,已被确认与慢性胃炎、消化性溃疡、胃黏膜相关淋巴样组织淋巴瘤和胃癌密切相关.根除hp可以减少37％人群胃癌的发生及阻止无癌前病变人群胃癌的发生[1].目前国内推荐的标准三联疗法hp根除率均在80％以下.如何避免和克服耐药,成功根除hp成为当前临床工作较为棘手的问题.微生态疗法为hp相关疾病的防治提供了新的思路.大量研究结果提示,益生菌对hp具有间接或直接的抑制作用[2].本研究的目的在于评价益生菌联合三联疗法7d或10 d疗法对hp根除的疗效.     

益生菌治疗肠易激综合征的可能机制

肠易激综合征（irritable bowel syndrome．IBS）是常见的功能性胃肠病的一种,以反复发作的腹痛或腹部不适为特点,伴有大便性状和排便习惯的改变。据文献报道,IBS在欧美的发病率达到了20％,     

兔髓核细胞体外最佳培养条件的探索

[目的]探索髓核细胞培养的最佳条件，为进一步作为椎间盘组织工程种子细胞提供可能。[方法]以DMEM／F12(1：1，添加FBS10％，pH7．2)作为基础培养液，通过依次改变培养液pH、胎牛血清浓度、抗坏血酸浓度、培养温度、CO2浓度以及培养板处理，计数10d克隆形成数，检测细胞生长代谢，确定最佳培养条件。[结果](1)髓核细胞不易贴壁生长，在未经处理的培养板中进行单层培养细胞增殖能力较弱，克隆形成少；而铺被多聚赖氨酸的培养板中培养可以较快贴壁并形成相对较多的克隆；(2)生长培养液在DMEM／F12(1：1)，pH7．0、胎牛血清浓度15％、抗坏血酸30μg／ml、温度37℃以及CO2浓度为5％时，髓核细胞生长良好，Ⅱ型胶原和聚集蛋白聚糖(aggrecan)mRNA表达增高。[结论]在最佳培养条件下，髓核细胞生长状态良好，为椎间盘组织工程髓核细胞的培养奠定了基础。     

植物乳杆菌HO-69产抗菌肽的培养条件

对L．plantarum HO-69产抗菌肽的营养和培养条件进行了研究。实验结果袁明，MRS培养基是其产抗菌肽的最适培养体系。以碳源、氮源、缓冲盐等为研究因子，对MRS设计部分因子重复实验，采用回归分析确定K2HPO4和牛肉膏为抗菌肽产生的显著影响因子，综合考虑抗菌肽活力与纯化的难度，确定K2HPO4与牛肉膏的质量浓度分别为12g／L与16g／L，蛋白胨质量浓度为6g／L。响应面分析确定HO-69产抗菌肽的最适培养条件为：起始pH值6．61，36．12℃发酵13．87h，在此条件下发酵液的效价由80AU／mL提高到320AU／mL，抗菌肽的产量增加为原来的4倍。     

益生菌应用于肝移植术后患者的临床意义

目的：探讨益生菌应用于肝移植术后患者的临床价值。方法：将肝移植术后患者55例分为益生菌组和对照组,患者术后均给予免疫抑制剂,而益生菌组加服双歧杆菌四联活菌片1周（思连康,1.5 g,3次/d）。于术前及术后第2,5,8天检测患者血清胆红素（TBIL）,丙氨酸转氨酶（ALT）,血清清蛋白（ALB）,前清蛋白（PA）水平,并观察和记录患者术后感染及胃肠道不良反应的发生情况。结果：TBIL和ALT水平两组在术前及术后第2天均无统计学差异（均P>0.05）,但益生菌组TBIL和ALT水平分别在术后第8,5天开始明显低于对照组（均P<0.05）;ALB和PA水平两组在术前及术后第2天均无统计学差异（均P>0.05）,而术后第5,8天益生菌组的两者水平均明显高于对照组（均P<0.05）;术后1周内益生菌组与对照组的感染率及不良反应发生率分别为41.38%和69.23%（P=0.038）,17.24%和42.31%（P=0.041）。结论：肝移植术后应用益生菌可以改善机体的营养状况,促进肝功能恢复,并减轻炎症反应、降低术后感染及胃肠道不良反应的发生率。     

益生菌制品消费水平对维持性血液透析患者焦虑及抑郁的影响

目的探讨维持性血液透析(maintenance hemodialysis,MHD)患者益生菌制品消费情况对患者抑郁、焦虑状态的影响。方法纳入MHD患者102例,根据过去半年益生菌制品包括酸奶和肠道微生物制剂消费情况进行分组,运用焦虑自评量表(self-rating anxiety scale,SAS)、抑郁自评量表(self-rating depression scale,SDS)比较不同组间抑郁、焦虑状态的差异。结果纳入的患者中,酸奶高消费者50例(49.02%),肠道微生物制剂高消费者29例(28.43%),益生菌制品高消费者79例(77.45%);酸奶高消费组患者SAS、SDS评分低于酸奶低消费组,差异有统计学意义(P<0.05);肠道微生物制剂高消费组SAS、SDS评分均低于肠道微生物制剂低消费组,差异有统计学意义(P<0.05);益生菌制品高消费组SAS、SDS评分均低于益生菌制品低消费组,差异有统计学意义(P<0.05);酸奶高消费组SAS、SDS评分均高于肠道微生物制剂高消费组,差异有统计学意义(P<0.05)。结论益生菌制品消费水平越高,MHD患者的SAS、SDS评分越低,合理补充益生菌制品可能可以改善MHD患者焦虑、抑郁状态。益生菌制品对MHD患者情绪影响值得临床关注及进一步探讨。     

清热解毒方与益生菌联合治疗大鼠脓毒症

目的：观察清热解毒方和益生菌联合治疗脓毒症模型大鼠的作用。方法：采用盲肠结扎穿孔法制作大鼠脓毒症模型,健康Wistar大鼠140只随机分为假手术组、脓毒症模型组、抗生素组、益生菌组、清热解毒方中药组及清热解毒方和益生菌联合治疗组,对大鼠腹水和血标本进行细菌培养鉴定及实时定量PCR检测。结果：与假手术组比较,脓毒症模型组大鼠腹水和血液中细菌检出率明显升高（P＜0.05）,其腹水中大肠杆菌检出率100%、金黄色葡萄球菌检出率63.6%,血液中大肠杆菌检出率94.7%、金黄色葡萄球菌检出率21.1%。与模型组比较,单纯中药组和单纯益生菌组治疗后细菌检出率和数量均有所降低（P＜0.05）。与单纯用药组比较,联合治疗组上述细菌检出率和数量进一步降低（均P＜0.05）。结论：脓毒症模型大鼠主要致病菌为大肠杆菌和金黄色葡萄球菌,清热解毒方和益生菌联合使用可协同抑制其主要致病菌。     

The ameliorative effects of Lactobacillus coagulans and Lactobacillus casei probiotics on CCl4-induced testicular toxicity based on biochemical,histological and molecular analyses in rat

Probiotics are commonly present in foods and role as dietary adjuncts and alternatives to pharmacological products in many medical fields. The aim of this study was to investigate the effects of Lactobacillus coagulans and Lactobacillus casei probiotics on carbon tetrachloride (CCl₄)-induced reproductive injury and sperm toxicity in rats. Thirty-two Wistar rats were divided into four groups as follows: sham, CCl₄ (2 ml/kg), L. casei probiotic + 2 ml/kg CCl₄ and L. coagulans probiotic + 2 ml/kg CCl₄. On the 36th day after the intervention, serum levels of follicle-stimulating hormone (FSH), luteinising hormone (LH) and total testosterone (T), as well as catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities, were measured. Testicular malondialdehyde (MDA) level, the expressions of apoptosis-related genes (Bcl-2 and Bax), Bax/Bcl-2 ratio, histomorphometric indices such as tubular differentiation index (TDI), repopulation index (RI), spermiogenesis index (SPI) and sperm parameters were evaluated. L. casei and L. coagulans probiotics improved the levels of reproductive hormones and antioxidant capacity in rats. Both the probiotics, especially L. casei, increased the rate spermatogenesis which accompanied with significant increments in testicular TDI, RI and SPI. Furthermore, both probiotics down-regulated Bax and up-regulated Bcl-2, following by decreased Bax/Bcl-2 ratio. Our key findings indicated that L. casei and L. coagulans have protective effects against CCl₄-induced testicular toxicity.     

乳酸杆菌冻干发酵剂

以乳酸杆菌为菌种，进行单因素和响应面法实验优化得到制备冻干发酵剂的复合保护剂为：脱脂乳粉9．7g／dL，半胱氨酸0．03mol／L，谷氨酸钠3．2g／dL，所得冻干发酵剂存活率为89．5%。且该冻干发酵剂于4℃保存6个月后的存活率为66．4％。     

Effect of Lactobacillus casei on a novel murine model of abdominal sepsis

BACKGROUND: Lactobacilli are one of the probiotics and there is strong evidence for the efficacy of lactobacilli administration as a nonspecific immunostimulant in increasing host defense mechanisms. The present study tested the hypothesis that heat-killed Lactobacillus casei (LC9018) has a protective activity against fecal peritonitis. METHODS: Cecal ligation and tip resection (CLTR) induced fecal peritonitis was developed as a novel mouse model of abdominal sepsis, and the effects of LC9018 pretreatment on survival after CLTR, of the peritoneal exudate cells before or after CLTR and of bacterial growth in the peritoneal cavity after CLTR were investigated. RESULTS: Mortality after CLTR varied directly with the length of the opened bowel. To obtain a sublethal experimental group, the length of the opened bowel was fixed at 4 mm, where mortality was 87%, for further experiments. Survival of mice after CLTR was augmented in mice that had been pretreated intraperitoneally (ip) with LC9018 24 h previously. Viable bacterial growth in the peritoneal cavity was markedly inhibited in LC9018-pretreated mice. Peritoneal exudate cell accumulation observed 24 h after ip injection of LC9018 was significantly enhanced, suggesting that augmentation of the resistance of mice to CLTR was caused especially by the induction of polymorphonuclear cells. CONCLUSIONS: CLTR may provide a reproducible and simple murine model of bacterial sepsis and pretreatment with LC9018 developed a protective activity against CLTR surgery.     

Lactobacillus Peritonitis: A Rare Cause of Peritonitis in Peritoneal Dialysis Patients

Lactobacilli are part of the normal gastrointestinal and female genitourinary flora in humans and they are seldom pathogenic and rarely cause human disease. In the literature, Lactobacillus peritonitis was most common in immunocompromised patients, including patients under chronic peritoneal dialysis. We also suspect that the presence of Lactobacillus spp. in the peritoneal fluid might indicate the leakage of normal flora from a perforated intraabdominal hollow organ. To access the versatile clinical pictures of Lactobacillus peritonitis, this investigation retrospectively reviewed the medical records for Lactobacillus spp. isolated from peritoneal fluid from July 1998 to January 2002 at Chang‐Gung Memorial Hospital, Taipei, Taiwan. A total of 10 patients were enrolled in the study. Six of these 10 patients had concomitant intraabdominal hollow organ perforation, and peritoneal fluid cultures in these six patients also contained bacteria other than Lactobacillus spp. All six patients had recently experienced either abdominal surgery or blunt abdominal trauma. The remaining four patients who had not undergone surgery had decompensated liver cirrhosis with ascites and spontaneous bacterial peritonitis. The results suggested that the presence of Lactobacillus spp. in the peritoneal fluid other than immunocompromised patients should raise the suspicion of hollow organ perforation in patients with recent abdominal surgery or blunt abdominal trauma.     

乳酸杆菌肽聚糖的分离鉴定及其免疫活性

研究了乳酸杆菌细胞壁肽聚糖的分离、鉴定及其对肽聚糖生物活性的影响.实验获得了SDS处理粗细胞壁的适宜条件为SDS质量浓度8g/dL,处理时间为沸水浴10min.SDS结合胰蛋白酶和TCA处理,可有效去除乳酸杆菌中的非共价结合蛋白质及共价结合蛋白质.经化学分析鉴定,所提取的乳酸杆菌成分为肽聚糖,肽聚糖中丙氨酸、天冬氨酸、谷氨酸和赖氨酸浓度分别为1.181,0.943,0.770和0.456mmol/g,是肽聚糖中的组分氨基酸.SDS结合TCA处理,方能较有效地去除DNA.TCA处理是去除细胞壁磷壁酸、脂磷壁酸的有效方法.此外,小白鼠腹腔巨噬细胞的吞噬实验、C3b受体实验以及血清溶菌酶活力实验证实,所分离纯化肽聚糖的免疫学活性未受影响.     

The effect of Lactobacillus bacteria supplement on sepsis and its complications in patients with acute burns

Sepsis as a result of bacterial translocation from the gastrointestinal tract (GIT) is a known associate of morbidity and mortality in patients with severe burns. This translocation is influenced by the GIT flora. Oral consumption of Lactobacillus bacteria was previously shown to reduce translocation. We conducted a retrospective cohort study on a series of 56 patients with burns admitted to Soroka University Medical Center in Beer-Sheva, Israel. Those 56 patients included 28 who were given lactobacillus supplements and 28 who were not. The parameters that were compared between the groups evaluated the level of sepsis and its complications. The parameters of morbidity during hospitalization were significantly higher in the treatment group; however, their mortality was lower. That difference in mortality between the groups was not significant as a whole (p=0.071), but it was significant in the subgroup analysis of 41-70% total body surface area burned. In that subgroup there were zero cases of death in the treatment group versus five cases in the control group (p=0.005). Our findings suggest that in acute burns, lactobacillus bacteria food additives may be clinically beneficial in patients with total burned body surface area of 41-70%.     

乳酸菌SK1.002产L-阿拉伯糖异构酶培养基优化及发酵条件

L-阿拉伯糖异构酶能将D-半乳糖异构成D-塔格糖。通过单因素试验和快速登高法对乳酸菌SK1.002产L-阿拉伯糖异构酶的培养基进行优化,确定发酵优化条件为（组分g/L）：麦芽糊精28,酵母膏10,玉米粉浆22,无水乙酸钠10,K3PO40.2,NaCl 0.01,FeSO4.7H2O 0.01,Mg-SO4.7H2O 0.2,MnSO4.2H2O 0.05,L-阿拉伯糖2.5。发酵初始pH 8.4,培养温度37℃,接种体积分数3%,培养时间12 h。在此发酵条件下,酶活达到了7.28 U/mL。     

麦汁的生物酸化技术

作者研究了以德氏乳酸杆菌为生产茵种，以未添加酒花的麦汁作为培养基生产乳酸的主汁生物酸化技术，用于调节啤酒生产过程pH值。确定5^#乳酸菌的最适生长温度为42℃，最佳生物酸化麦汁质量分数为14％。研究表明，液体培养基的组成对该菌产乳酸有较大的影响。生物酸化麦汁调酸比约为1.40左右，不会对成品啤酒风味产生太大的影响。     

定期局部应用雌激素联合乳杆菌胶囊治疗老年萎缩性阴道炎的效果分析

目的 分析定期局部应用雌激素联合乳杆菌胶囊治疗老年萎缩性阴道炎的效果。方法 选取本院2021年5月-2022年5月收治的84例老年萎缩性阴道炎患者为研究对象,按照随机数字表法将其分为对照组和观察组,每组42例。对照组接受乳酸杆菌胶囊治疗,观察组采用雌激素联合乳酸杆菌胶囊治疗,比较两组临床疗效、满意度、生活质量、负面情绪、阴道pH值及不良反应发生情况。结果 观察组治疗总有效率高于对照组（P＜0.05）;观察组满意度高于对照组（P＜0.05）;观察组SF-36评分、阴道pH值、SAS、SDS评分均优于对照组（P＜0.05）;观察组不良反应总发生率低于对照组（P＜0.05）。结论 定期局部应用雌激素和乳酸杆菌胶囊可有效治疗老年萎缩性阴道炎,可提高患者生活质量与满意度,促进症状改善,且安全性较高。