Effect of the gastrointestinal environment on pH homeostasis of Lactobacillus plantarum and Lactobacillus brevis cells as measured by real-time fluorescence ratio-imaging microscopy

In the present work, an in vitro model of the gastrointestinal tract (GIT) was developed to obtain real-time observations of the pH homeostasis of single cells of probiotic Lactobacillus spp. strains as a measure of their physiological state. Changes in the intracellular pH (pH_i) were determined using fluorescence ratio imaging microscopy (FRIM) for potential probiotic strains of Lactobacillus plantarum UFLA CH3 and Lactobacillus brevis UFLA FFC199. Heterogeneous populations were observed, with pH_i values ranging from 6.5 to 7.5, 3.5 to 5.6 and 6.5 to 8.0 or higher during passage of saliva (pH 6.4), gastric (pH 3.5) and intestinal juices (pH 6.4), respectively. When nutrients were added to gastric juice, the isolate L. brevis significantly decreased its pH_i closer to the extracellular pH (pH_ex) than in gastric juice without nutrients. This was not the case for L. plantarum.This study is the first to produce an in vitro GIT model enabling real-time monitoring of pH homeostasis of single cells in response to the wide range of pH_ex of the GIT. Furthermore, it was possible to observe the heterogeneous response of single cells. The technique can be used to determine the survival and physiological conditions of potential probiotics and other microorganisms during passage through the GIT.     

根痛平胶囊和根痛平片中川续断皂苷Ⅵ的含量测定方法研究及续断的投料情况分析

目的 建立根痛平胶囊和根痛平片中川续断皂苷Ⅵ的含量测定方法,监测生产企业续断投料情况。方法 采用高效液相色谱(HPLC)法,Agilent Zorbax SB-C₁₈色谱柱(4.6 mm×250 mm,5 μm),以乙腈-水（27:73）为流动相等度洗脱,流速1.0 mL·min-1,检测波长为212 nm。结果 川续断皂苷Ⅵ在30~280 μg·mL^-1的范围内呈良好的线性关系,回归方程为Y = 2 029.357 0 X + 3.688 5 ,R² = 0.999 7,平均回收率为96.08%,RSD为1.01%。结论 该法简单准确,可用于测定根痛平胶囊和根痛平片中川续断皂苷Ⅵ的含量,为根痛平系列制剂的质量标准提高提供理论依据;制定了川续断皂苷Ⅵ含量参考限度值,考察企业续断的投料情况,提示个别企业未投料或投料用续断质量差。     

胸椎旁神经阻滞与硬膜外麻醉对非气管插管肺叶切除术患者镇痛和免疫的影响*

目的?比较胸椎旁神经阻滞（TPVB）和硬膜外阻滞（EB）对非气管插管肺叶切除术患者镇痛和免疫的影响。方法?选取2019年1月—2019年6月在青岛大学附属青岛市市立医院东院区行胸腔镜下肺叶切除术的120例非小细胞肺癌患者,用随机数字表法分为TPVB组、EB组和全身麻醉（GA）组。比较各组患者围手术期镇痛和芬太尼追加量情况、细胞免疫及体液免疫水平。结果?3组术后1?h、2?h、6?h、12?h、24?h和48?h VAS评分在不同时间、不同组间及变化趋势上有差异（P?<0.05）。3组芬太尼追加量比较,差异有统计学意义（P?<0.05）,GA组高于TPVB组、EB组（P?<0.05）。3组术后第1天和第3天CD3+CD4+、CD3+CD8+水平在不同时间、不同组间有差异（P?<0.05）,变化趋势无差异（P?>0.05）。3组术后第1天和第3天CD3-CD16+CD56+、CD19+水平在不同时间、不同组间及变化趋势上无差异（P?>0.05）。3组术后第1天和第3天IgG、IgM、IgA在不同时间有差异（P?<0.05）,不同组间及变化趋势上无差异（P?>0.05）。与EB组比较,TPVB组患者围手术期低血压发生率降低（P?<0.05）。结论?对于非气管插管肺叶切除术患者,TPVB和EB均能提供有效的围手术期镇痛;同时TPVB术后镇痛时间更持久,术后T淋巴细胞免疫抑制减轻。     

白细胞介素18单核苷酸多态性与73例喉鳞状细胞癌易感性的关联性

目的 探讨白细胞介素18(IL-18)启动子位点-137G/C和-607C/A基因多态性与山东地区汉族人群喉鳞状细胞癌(LSCC)易感性的关系。 方法 随机选取2017年5月至2019年10月就诊于临沂市人民医院的喉鳞状细胞癌患者73例为病例组,选取同期查体的健康志愿者73人为健康对照组。收集研究对象一般临床资料及外周静脉血2 mL,提取全血DNA,采用sanger测序法进行基因测序及SNP分型。所有定量资料采用Students-t检验分析,定性资料采用Pearson χ2检验或Fisher精确检验,应用卡方拟合优度检验分析健康对照组是否符合Hardy-Weinberg平衡定律,采用二元Logistic回归分析计算两种单核苷酸多态性(SNP)位点以及各风险因素(年龄、吸烟、饮酒)与LSCC发病风险的关联性。 结果 与健康对照组相比,病例组中-137G/C位点变异基因型GC的基因型频率升高(34.2% vs 19.2%, P=0.040),而-607C/A 位点各基因型频率的分布在病例组和健康对照组之间差异无统计学意义(P>0.05)。吸烟及饮酒均会升高LSCC的发病风险(OR分别为5.710和3.116)。对基因型、年龄组、吸烟史及饮酒史4种风险因素进行调整后,吸烟者与-137G/C基因型携带者LSCC发病风险的OR值分别为4.967(95%CI=2.185~11.289, P<0.001)和2.971(95%CI=1.243~7.101, P=0.014),差异有统计学意义。 结论 IL-18启动子位点-137G/C而非-607C/A的多态性与LSCC的患病风险升高有关联性。     

天麻指纹图谱的研究进展

天麻是我国名贵的中草药,随着近年来其商品的多样化,掺伪、假冒品不断增多,对其进行更全面、系统的质量控制尤为重要。目前,指纹图谱已成为国际公认的控制中药质量的有效手段之一,主要的研究方法有液相色谱法、薄层色谱法、毛细管电泳色谱、红外光谱、核磁共振、X射线粉末衍射、分子生物学指纹图谱和电化学指纹图谱等,本文对近年来天麻指纹图谱的研究情况进行了归纳总结,为进一步建立该药的质量控制提供了方法和依据。     

987例乙型肝炎患者HBV-DNA P区多位点耐药突变分析及临床意义

目的分析乙型肝炎患者HBV-DNA P区基因突变情况、用药史及其临床意义。方法采用焦磷酸测序法,对368例经核苷(酸)类似物(NAs)治疗1年以上(用药组)及619例未用抗病毒药物治疗(未用药组)的乙型肝炎患者HBV-DNA P基因区9个NAs相关耐药突变位点进行检测,回顾性分析不同NAs耐药的突变形式,分析NAs治疗前HBV逆转录酶(RT)基因自然变异的发生率。结果 368例中94例出现基因耐药变异,变异模式以经典突变L180M、M204V/I、A181V/T、N236T突变为主,其次为V173L、S202G/S、T184L、I169T突变,M204V多以联合L180M突变的形式存在,其中235例服用单种NAs有48例发生变异,多位点(3个及以上耐药位点)突变5例(10.42%),133例服用多种NAs有46例发生变异,多位点突变19例(41.30%),多药使用者发生多位点变异率明显高于单药使用者(χ2=8.38,P0.05)。368例患者中多位点变异组(A组)与少于三个位点变异组(B组)的年龄、HBVDNA、ALT无明显差异。619例中51例出现基因耐药变异(8.24%)。结论 HBV感染者存在少量自然变异位点。长期应用NAs可筛选出HBV-DNA P基因区的相关耐药变异且耐药变异复杂,服用多种NAs(特别是序贯治疗)容易发生基因耐药变异且易发生多位点变异。长期应用NAs的乙型肝炎患者需定期检测基因耐药情况并及时干预,避免临床反跳的发生。     

偏侧咀嚼对大鼠颞下颌关节滑膜VEGF表达影响的研究

目的:研究偏侧咀嚼对大鼠颞下颌关节(TMJ)滑膜血管内皮细胞生长因子(VEGF)表达的影响。方法:30只大白鼠随机分为6组,实验组间断磨除右侧上、下颌磨牙牙冠至龈下,对照组未做处理,饲养条件相同。实验1组磨除牙冠后4周末、实验2、3组磨除牙冠后10、16周末处死动物,取其双侧TMJ切片免疫组化检测。结果:实验1、2、3组双侧颞下颌关节滑膜VEGF表达较对照组增强。结论:偏侧咀嚼可引起颞下颌关节滑膜损伤,VEGF参与了其病理过程。     

Elevated Th17 cells are accompanied by FoxP3+ Treg cells decrease in patients with lupus nephritis

To investigate the variations of T-helper 17 (Th17) and regulatory T (Treg) cells in patients with lupus nephritis (LN), a total of 60 systemic lupus erythematosus patients and 28 healthy controls (HCs) were enrolled. The frequency of Th17 cells and Treg cells in peripheral blood mononuclear cells (PBMCs) was evaluated by flow cytometric analysis. The serum concentrations of interleukin-17 (IL-17) and transforming growth factor-beta 1 (TGF-β1) were measured by enzyme-linked immunosorbent assay (ELISA). The results demonstrated in LN patients a significant decrease in the frequency of CD4+CD25^high and CD4+CD25+FoxP3+ T cells and a significant increase in the frequency of Th17 cells in peripheral blood, and the ratio of Th17 to Treg cell frequency was significantly increased along with increased SLEDAI scores. LN patients had a lower percentage and expression of FoxP3 in CD4+CD25^high T cells than SLE patients without nephritis. The concentration of TGF-β1 was found decreased in SLE patients compared with that from healthy controls, though no significant difference was found between LN patients and SLE patients without nephritis. The expression of IL-17 levels in LN patients exhibited a significant increase compared with patients without nephritis and healthy controls. Based on our results, the significantly elevated Th17 cells are accompanied by FoxP3+ Treg cells decrease in lupus nephritis, suggesting that Th17/Treg functional imbalance may be involved in the pathogenesis of renal damage in SLE patients.     

Association of TOMM40 Polymorphisms with Late-Onset Alzheimer’s Disease in a Northern Han Chinese Population

Mitochondrial dysfunction is an early defect in the pathogenesis of late-onset Alzheimer’s disease (LOAD). One interesting candidate gene for mitochondrial dysfunction in LOAD is the translocase of outer mitochondrial membrane 40 homolog (TOMM40) gene. Several single nucleotide polymorphisms (SNPs) within TOMM40 have been shown to affect susceptibility to LOAD in Caucasians, while there are no studies on the association of the polymorphisms with LOAD risk in Han Chinese. Here, the association of TOMM40 polymorphisms in LOAD was investigated in a large Northern Han Chinese cohort consisting of 1,578 individuals. Both allelic and genotypic associations of three SNPs (rs157580, rs2075650, and rs11556505) with LOAD risk were observed in the total sample as well as in the non- APOE ε4 carriers. For rs1160985, the allele and genotype frequencies differed significantly only in APOE ε4 carriers. After adjustment for age, gender, and APOE ε4 status, the association remained statistically significant only for the rs157580 but not for rs2075650 and rs11556505. In contrast, the rs1160985 exhibited significant risk effect after adjustment. In addition, haplotype analysis confirmed that the haplotypes derived from SNPs in rs2075650, rs11556505, and rs1160985 were associated with either risk or protective effects. In summary, our findings suggest that the TOMM40 polymorphisms may play a role in the pathogenesis of LOAD in Han Chinese.