Selection of Gut-Resistant Bacteria and Construction of Microbial Consortia for Improving Gluten Digestion under Simulated Gastrointestinal Conditions

This work aimed to define the microbial consortia that are able to digest gluten into non-toxic and non-immunogenic peptides in the human gastrointestinal tract. Methods: 131 out of 504 tested Bacillus and lactic acid bacteria, specifically Bacillus (64), lactobacilli (63), Pediococcus (1), and Weissella (3), showed strong gastrointestinal resistance and were selected for their PepN, PepI, PepX, PepO, and PepP activities toward synthetic substrates. Based on multivariate analysis, 24 strains were clearly distinct from the other tested strains based on having the highest enzymatic activities. As estimated by RP-HPLC and nano-ESI–MS/MS, 6 cytoplasmic extracts out of 24 selected strains showed the ability to hydrolyze immunogenic epitopes, specifically 57–68 of α9-gliadin, 62–75 of A-gliadin, 134–153 of γ-gliadin, and 57–89 (33-mer) of α2-gliadin. Live and lysed cells of selected strains were combined into different microbial consortia for hydrolyzing gluten under gastrointestinal conditions. Commercial proteolytic enzymes (Aspergillus oryzae E1, Aspergillus niger E2, Bacillus subtilis Veron HPP, and Veron PS proteases) were also added to each microbial consortium. Consortium activity was evaluated by ELISA tests, RP-HPLC-nano-ESI–MS/MS, and duodenal explants from celiac disease patients. Results: two microbial consortia (Consortium 4: Lactiplantibacillus (Lp.) plantarum DSM33363 and DSM33364, Lacticaseibacillus (Lc.) paracasei DSM33373, Bacillus subtilis DSM33298, and Bacillus pumilus DSM33301; and Consortium 16: Lp. plantarum DSM33363 and DSM33364, Lc. paracasei DSM33373, Limosilactobacillus reuteri DSM33374, Bacillus megaterium DSM33300, B. pumilus DSM33297 and DSM33355), containing commercial enzymes, were able to hydrolyze gluten to non-toxic and non-immunogenic peptides under gastrointestinal conditions. Conclusions: the results of this study provide evidence that selected microbial consortia could potentially improve the digestion of gluten in gluten-sensitive patients by hydrolyzing the immunogenic peptides during gastrointestinal digestion.     

聚合酶链反应检验法在阴道炎患者阴道细菌检查中的应用价值

目的探讨聚合酶链反应检验法在阴道炎患者阴道细菌检查中的应用价值。方法选取2019年6月~2020年6月于我院行细菌检查的58例阴道炎患者为研究对象,所有患者均采用阴道分泌物细菌培养、聚合酶链式反应(PCR)检验,比较两种检验方法阳性检出率。结果PCR检验阳性检出率(89.66%)高于细菌培养法(70.69%),差异有统计学意义(P<0.05)。结论PCR检验法应于阴道炎患者阴道细菌检查中,阳性检出率高,可为阴道炎的诊断及治疗提供较准确的科学依据,具有较高的临床应用价值。     

Fecal Excretion of Mycobacterium leprae,Burkina Faso

Mycobacterium leprae was detected by optical microscopy, fluorescent in situ hybridization, and molecular detection in feces collected for the diagnosis of Entamoeba coli enteritis in a leprosy patient in Burkina Faso. This observation raises questions about the role of fecal excretion of M. leprae in the natural history and diagnosis of leprosy.     

模拟复用的医用外科口罩清洗和消毒后主要性能指标变化

目的 医用外科口罩模拟复用清洗和消毒后主要性能指标变化。方法 以颗粒过滤效率（PFE）、细菌过滤效率（BFE）作为医用外科口罩的关键性能指标,对不同佩戴时间、不同干燥方式、不同洗消方式等试验条件进行验证。结果 试验后的医用外科口罩关键性能指标能够符合标准要求。结论 医用外科口罩采用清水洗涤、56℃热水浸泡和100℃沸水浸泡处理后主要性能指标未见明显下降。     

阴道微环境失衡与高危型人乳头瘤病毒持续感染

目的探讨阴道微生态失衡与高危型人乳头瘤病毒(HR-HPV)持续感染的关系。 方法选择2014年1月至2015年12月,在四川省科学城医院妇科诊断为HR-HPV感染的240例患者为研究对象,年龄为19~73岁。根据HR-HPV感染状态,将其分为研究组(n＝121,HR-HPV持续感染者)和对照组(n＝119,HR-HPV复查呈阴性者)。采用成组t检验,对2组患者的体重、人体质量指数(BMI)、孕次、产次等进行统计学分析。采用χ²检验,对2组患者的细菌性阴道病(BV)、外阴阴道假丝酵母菌病(VVC)、滴虫性阴道炎(TV)、需氧菌性阴道炎(AV)发生率等进行统计学分析。本研究遵循的程序符合2013年修订的《世界医学协会赫尔辛基宣言》要求。 结果①2组受试者体重、BMI、孕次和产次等一般临床资料比较,差异均无统计学意义(P>0.05)。②研究组患者BV、VVC检出率分别为43.0%(52/121)、38.0%(46/121),明显高于对照组的20.2%(24/119)、15.1%(18/119),2组比较,差异均有统计学意义(χ²＝14.422、16.075,P＝0.002、<0.001)。研究组和对照组患者的TV、AV和混合性感染患者检出率比较,差异均无统计学意义(P>0.05)。 结论BV、VVC与HR-HPV持续性感染密切相关,改善阴道微生态环境可能有助于降低HR-HPV的发生率。     

Genomic Characterization of hlyF-positive Shiga Toxin–Producing Escherichia coli,Italy and the Netherlands, 2000–2019

Shiga toxin–producing Escherichia coli (STEC) O80:H2 has emerged in Europe as a cause of hemolytic uremic syndrome associated with bacteremia. STEC O80:H2 harbors the mosaic plasmid pR444_A, which combines several virulence genes, including hlyF and antimicrobial resistance genes. pR444_A is found in some extraintestinal pathogenic E. coli (ExPEC) strains. We identified and characterized 53 STEC strains with ExPEC-associated virulence genes isolated in Italy and the Netherlands during 2000–2019. The isolates belong to 2 major populations: 1 belongs to sequence type 301 and harbors diverse stx₂ subtypes, the intimin variant eae-ξ, and pO157-like and pR444_A plasmids; 1 consists of strains belonging to various sequence types, some of which lack the pO157 plasmid, the locus of enterocyte effacement, and the antimicrobial resistance–encoding region. Our results showed that STEC strains harboring ExPEC-associated virulence genes can include multiple serotypes and that the pR444_A plasmid can be acquired and mobilized by STEC strains.     

我国常见细菌耐药趋势预测研究:基于灰色GM (1,1)模型

 目的 通过对耐甲氧西林金黄色葡萄球菌（MRSA）、耐碳青霉烯类铜绿假单胞菌（CRPA）、耐碳青霉烯类鲍曼不动杆菌（CRAB）、耐第三代头孢菌素的大肠埃希菌（3GCR-E.coli）、耐第三代头孢菌素的肺炎克雷伯菌（3GCR-KP）等细菌耐药数据构建灰色预测模型,分析细菌耐药特征的变化趋势,探讨灰色预测模型在细菌耐药领域的应用价值。方法 采用2014-2018年全国细菌耐药监测报告中MRSA、CRPA和CRAB、3GCR-E.coli、3GCR-KP等耐药率数据构建灰色预测GM （1,1）模型。用后验差比C值和小误差概率P值评估模型精度,用相对误差和级比偏差评估模型拟合效果,并用2019-2020年数据对模型预测效果进行验证。最终根据模型对2021-2023年的耐药率进行预测。结果 本研究构建的GM （1,1）模型对MRSA、CRPA、CRAB、3GCR-E.coli和3GCR-KP等细菌耐药率预测效果较好,根据该模型预测到2023年其耐药率分别可降低至23.9%、15.2%、50.2%、43.8%、26.1%。结论 全国针对细菌耐药情况采取的控制措施取得明显成效,GM （1,1）模型对细菌耐药率预测效果较好,可在细菌耐药管理领域推广应用。     

PCT与SDI比值对重症细菌性肺炎患者短期预后的预测价值

 目的 探讨血清降钙素原(PCT)与肺泡灌洗液辛普森菌群多样性指数(SDI)比值对重症监护病房(ICU)内细菌性肺炎患者短期预后的预测价值。方法 回顾性调查扬州大学附属医院ICU 2019年10月—2021年7月选择肺泡灌洗液宏基因组二代测序(mNGS)技术的56例细菌性肺炎患者病历资料,依据其入ICU 24 h内急性生理学与慢性健康状况评分Ⅱ(APACHE-Ⅱ)分为非危重症组21例和危重症组35例。以细菌性肺炎造成死亡为终点事件,记录28天转归,并将患者分为生存组38例和死亡组18例。对各组患者的SDI、PCT、C-反应蛋白(CRP)、PCT/SDI、CRP/SDI进行比较分析。结果 与非危重症组比较,危重症组患者血清PCT/SDI、PCT水平均升高,且呼吸机辅助通气时间更长,28天病死率更高(均P＜0.05);与存活组比较,死亡组患者SDI较低,PCT/SDI、PCT水平均较高(均P＜0.05);SDI与呼吸机辅助通气时间呈负相关(r值为-0.655,P＜0.001),PCT水平、PCT/SDI与呼吸机辅助通气时间呈正相关(r值分别为0.660、0.734,均P＜0.001)。受试者工作特征曲线(ROC曲线)显示,PCT/SDI预测患者28天死亡的ROC曲线下面积(AUC)为0.851,其次为PCT+SDI (0.845)、PCT (0.808)、SDI (0.785)、CRP/SDI (0.731),PCT/SDI的最佳截断值为11.56时预判患者28天死亡的灵敏度为89.5%,特异度为66.7%。Cox回归分析显示,PCT/SDI值高(HR=1.562,95%CI：1.271~1.920,P=0.031)、PCT水平高(HR=1.106,95%CI：1.021~1.198,P=0.024)是ICU细菌性肺炎患者死亡的独立危险因素。结论 PCT/SDI、PCT、PCT+SDI、SDI、CRP/SDI均可作为ICU细菌性肺炎患者短期预后的评估指标。与其他指标相比,PCT/SDI预测患者短期预后更有价值。     

新疆喀什市细菌性阴道病（BV）的调查

目的：了解细菌性阴道病（BV）在喀什市育齿妇女普查人群中的患病情况。方法：对822名妇科普查的妇女按Amsel等对细菌性阴道病（BV）诊断的4项标准进行检测及宫颈防癌涂片筛查。结果：细菌性阴道病（BV）在普查妇女中患病率为19.59%(161/822)。患者无主诉、无明显白带增多等症状占68.32%(110/161)。不孕症14例占8.69%(14/161),合并滴虫、霉菌性阴道炎出现阴道充血、瘙7痒等症状来就诊检查者51例。占40.37%（65/161）,有阴道上尾丝环史104例。结论：建议有条件的医院及计划生育指导所,在妇科生殖健康服务时,应将查BGV列为常规门诊检测项目。     

羊水葡萄糖及白介素-6在胎膜早破中的意义

目的 探讨胎膜早破患者羊水中葡萄及白介素－6(IL－6）含量与宫内感染的关系。方法 随机选择1994－1999年70例无妊娠并发症的胎膜早破的患者,在B超引导下,经腹行羊膜腔内穿刺,抽取羊水,用葡萄糖氧化酶方法测定水中葡萄糖,用酶联免疫方法测定IL－6含量。结果 胎膜早破并宫内感染者,羊水葡萄糖含量明显下降,IL－6含量则明显增高,当羊水葡萄糖含量≤0．55mmol/L,IL-6含量≥315ng/L时,预测宫内感染的敏感性100％,特异性＞80．00％。结论 胎膜早破患者,测定羊水葡萄糖及IL－6含量,能早期快速预测宫内感染。