半夏泻心汤对DSS诱导的溃疡性结肠炎小鼠肠道微生态及Th17/Treg细胞平衡的影响＊

目的 观察半夏泻心汤对饮用葡聚糖硫酸钠（DSS）诱导建立的溃疡性结肠炎（UC）模型小鼠Th17/Treg细胞平衡及肠道微生态的影响，从免疫失衡及菌群紊乱角度分析半夏泻心汤治疗UC的作用机制。方法 将40只C57BL/6J雄性小鼠随机分为空白组、模型组、阳性药（美沙拉嗪）组及半夏泻心汤组，除空白组外的各小鼠采用2.5% DSS水溶液自由饮用7天诱导建立UC小鼠模型。自实验第8日起，分别采用灭菌水、美沙拉嗪水溶液、半夏泻心汤水溶液给上述各组别小鼠灌胃。采用HE染色观察结肠组织病理学变化，ELISA方法检测组织炎症因子含量，流式细胞术检测Th17及Treg细胞的相对数量，16S rRNA测序技术进一步检测模型小鼠粪便中菌群的变化。结果 与空白组比较，模型组小鼠结肠损伤严重，血清炎症因子含量显著升高（P<0.01）；与模型组比较，中药干预组小鼠结肠损伤减轻，血清TNF-α、IFN-γ含量显著下降（P<0.05）。较空白组小鼠相比，模型组小鼠Th17细胞百分比显著升高（P<0.01），而Treg细胞百分比显著降低（P<0.01）。经半夏泻心汤干预后，其小鼠Th17细胞数量均显著降低（P<0.05），Treg细胞数量则显著升高（P<0.05）。同时，半夏泻心汤可升高门水平上Firmicutes、属水平上Lachnospiraceae_NK4A136_group、unclassified_f_Lachnospiraceae、Prevotellaceae_UCG-001、Alistipes；降低门水平上Actinobacteriota，科水平上Bifidobacteriaceae、Atopobiaceae、Clostridiaceae，属水平上Clostridium_sensu_stricto_1、Coriobacteriaceae_UCG-002、Bifidobacterium的相对丰度。结论 半夏泻心汤可使Treg细胞相对数量升高、Th17细胞相对数量降低，以维持Th17/Treg细胞动态平衡；同时可降低有害菌Clostridium_sensu_stricto_1、Coriobacteriaceae_UCG-002的相对丰度，升高产短链脂肪酸类菌Lachnospiraceae_NK4A136_group、Prevotellaceae_UCG-001、unclassified_f_Lachnospiraceae等菌属的相对丰度，调控肠道微生物结构及动态平衡，从而发挥治疗UC的作用。     

大黄附子汤对重症急性胰腺炎小鼠肠道微皱褶细胞变化的影响

目的研究大黄附子汤对重症急性胰腺炎(SAP)小鼠肠道微皱褶细胞(M细胞)变化的影响，为大黄附子汤临床防治SAP提供理论基础。 方法选取40只健康SPF级C57BL/6J雄性小鼠，随机分为4组(每组10只)：空白对照组、大黄附子汤对照组、SAP组、大黄附子汤治疗组，其中SAP组和大黄附子汤治疗组分别以3.5 g/kg剂量腹腔注射20% L-精氨酸，空白对照组和大黄附子汤对照组注射等剂量的等渗盐水；于造模后12、24、36 h，空白对照组和SAP组给予等渗盐水0.2 mL，大黄附子汤对照组和治疗组给予大黄附子汤0.2 mL灌肠，均于造模完成48 h后处死取材，使用ELISA检测血清淀粉酶、内毒素、IL-1β及TNF-α含量，取回肠及胰腺组织行HE染色、评分，免疫组化染色检测M细胞特异性蛋白GP2并评分。 结果(1)一般情况：对照组腹腔注射后小鼠一般情况好，精神状态良好，能正常进水，四肢活动自如，行动未受影响；SAP组小鼠腹腔注射后一般情况差，精神萎靡，反应迟钝，行动迟缓，呼吸急促，拱背收腹，饮水减少。(2)SAP组淀粉酶、内毒素、IL-1β及TNF-α水平较对照组明显升高(P<0.05)；与SAP组进行对比，大黄附子汤治疗组血清淀粉酶、内毒素、IL-1β及TNF-α水平出现显著下降(P<0.05)。(3)HE染色：SAP组胰腺及回肠组织坏死严重，可见大量白细胞浸润。大黄附子汤治疗组胰腺及回肠组织轻度坏死，可见少量中性粒细胞等白细胞浸润。(4)免疫组化染色：SAP组与对照组相比GP2表达降低(P<0.05)；相较于SAP组，大黄附子汤治疗组GP2的表达水平升高(P<0.05)。 结论大黄附子汤治疗可改善作为肠道免疫应答起始的M细胞数量与功能，增强肠道免疫应答，减轻肠免疫屏障损伤，减少内毒素入血，改善SAP症状。     

ECFS standards of care on CFTR-related disorders: Updated diagnostic criteria

This paper is the first in a series providing updated guidance on the definition, evaluation and management of people with a Cystic Fibrosis Transmembrane conductance Regulator (CFTR)-Related Disorder (CFTR-RD). The need for this update relates to more precise characterisation of CFTR gene variants and improved assessment of CFTR protein dysfunction. The exercise is co-ordinated by the European CF Society Standards of Care Committee and Diagnostic Network Working Group and involves stakeholder engagement. This first paper was produced by a core group using an extensive literature review and papers graded for their quality. Subsequent wider stakeholder agreement was achieved.The definition of a CFTR-RD remains “a clinical condition with evidence of CFTR protein dysfunction that does not fulfil the diagnostic criteria for CF”. Clearer guidance on CFTR dysfunction and relevant CFTR variants will be provided. Thresholds for clinical presentations are presented and the paradigm that pathobiological processes may be evident in more than one organ is agreed. In this paper we reflect on the early patient journey, highlighting that CF specialists as well as other relevant specialists should be involved in the care of people with a CFTR-RD.     

Comparison of the effectiveness of i-scan and conventional endoscopy in the detection of the endoscopic signs of atrophic gastritis: A clinical trial

Background and study aimsThis study aimed to determine whether the use of i-scan endoscopy provides additional benefits to conventional endoscopy in the diagnosis of gastric precancerous lesions.Patients and methodsA total of 120 patients with histologically-verified intestinal metaplasia (IM) or atrophic gastritis (AG) were prospectively evaluated by esophagogastroduodenoscopy. Endoscopic examinations were performed using i-scan and high-definition white-light endoscopy (HD-WLE). The diagnostic yields of both techniques and the number of targeted biopsies per patient were compared.ResultsA total of 318 suspicious lesions were detected in 108 patients with i-scan (n = 186) and 81 patients with HD-WLE (n = 132). The diagnostic yields of i-scan and HD-WLE were 81.6% (98/120) versus 77.5% (93/120), respectively (p > 0.05). When only targeted biopsies were taken into account, the diagnostic yields of i-scan and HD-WLE were 89.8% versus 65.4%, respectively (p < 0.05). The mean number of biopsies per patient for i-scan and HD-WLE were 3.27 (393/120) and 7.3 (882/120), respectively (p < 0.05). The mean endoscopic procedure times were 16 and 17 min for i-scan and HD-WLE, respectively (p > 0.05).ConclusionsAlthough targeted biopsies with i-scan were not found to be significantly superior to either targeted or random biopsies with HD-WLE, the number of biopsies required to confirm these lesions was much lower.     

通过宏基因组学研究肠道微生物对结直肠癌患者影响的最新进展

人类肠道微生物已越来越受到医学界的广泛关注,人类肠道微生物不仅在人类健康方面,而且在人类疾病的发生和发展方面都有很大的作用。人们不断发现微生物与癌症之间的联系,特别是肠道微生物群和肠道肿瘤之间的联系。宏基因组学作为微生物研究的一种重要研究方法,在微生物与结直肠癌研究中发挥越来越重要的作用。近年来,通过宏基因组学研究肠道微生物菌群的变化为结直肠癌的发生和发展提供了新的见解,并且强调了癌症微生物群中宿主-微生物和微生物间相互作用的重要性。本综述回顾了通过宏基因组学研究肠道微生物与结直肠癌之间的关系,希望为癌症预防、诊断和治疗提供新的机会。     

益生菌联合肠内营养对急性胰腺炎患者肠道菌群及外周血miR-155的影响

目的 探讨益生菌联合肠内营养对急性胰腺炎患者肠道菌群及外周血miR-155的影响。 方法 选取2016年6月至2018年12月河北医科大学第二医院消化内科收治的92例急性胰腺炎患者作为研究对象，根据随机数字表法分为观察组（n=46）和对照组（n=46），对照组在常规治疗基础上给予肠内营养治疗，观察组在对照组基础上给予双歧杆菌乳杆菌三联活菌片，观察两组患者治疗前后肠道菌群、炎性介质及血清miR-155变化，比较两组患者治疗后胃肠道功能恢复情况。结果 治疗后，观察组双歧杆菌、乳酸杆菌增加数量，肠球菌、大肠埃希菌减少数量，血清CRP、IL-6、IL-17、TNF-α、miR-155降低水平均显著高于对照组（P<0.05）；观察组患者胃肠生活质量量表（GIQLI）评分高于对照组，腹胀消失时间、腹痛消失时间、肠鸣音恢复时间、血清淀粉酶恢复正常时间均少于对照组（P<0.05）。结论 益生菌联合肠内营养治疗急性胰腺炎可调节肠道菌群平衡，降低血清miR-155水平，缓解炎性反应，促进肠道功能恢复。     

Human breast milk oligosaccharides attenuate necrotizing enterocolitis in rats by suppressing mast cell accumulation,DPPI activity and TLR4 expression in ileum tissue,and regulating mitochondrial damage of Caco-2 cells

Necrotizing enterocolitis (NEC), a devastating infant disease characterized by severe intestinal necrosis, its pathogenesis is poorly understood, but appears to be multifactorial and highly associated with immaturity of gastrointestinal tract and immature innate-immune system. Breast-milk is effective strategy to protect infants against NEC. This study is using a NEC rat model to investigate the pathological mechanism of NEC involved intestinal-damages, and the therapeutic mechanism of sialylated human milk oligosaccharides (SHMOs) on NEC rats; also using cell model to investigate the effects of SHMOs on colon-epithelial cells (Caco-2) in-vitro.Extraction and characterization of SHMOs from breast milk, establishment of a NEC rat model, histopathological analysis and mast cell accounting of the terminal ileum were taken; The levels of DPPI, TLR4, IL-6, TNF-α, MMP-2/9 and glutathione were measured using various methods. Caco-2 cells were pre-treated with SHMOs and cultured with LPS, histamine, chymase or DPPI, cell viabilities and mitochondrial membrane potential were examined; flow cytometry was used to detect cell cycle.The accumulation of mast cells was found in the ileum of NEC rats, but prohibited by SHMOs treatment; the increased levels of TLR4, DPPI, IL-6, TNF-α, MMP-2/9 in NEC ileum were suppressed by SHMOs in-vivo. SHMOs prevented Caco-2 cells from LPS, histamine, chymase induced damages by surviving cell viability, regulating G0/G1 and S phase in cell cycles, and increasing mitochondrial membrane potential. These findings provide a new insight into the pharmacological mechanism of SHMOs treatment for NEC and suggest that SHMOs needs well attention for therapeutic aims.     

Effect of different modeling time on intestinal bacteria in letrozole induced PCOS rats

Objective: The differences of ovarian morphology, reproductive hormones, glucose and lipid metabolism and intestinal bacteria in rats with polycystic ovary syndrome (PCOS) induced by triazole were compared. Method: Eighteen 21 SPF female SD rats were randomly divided into group A (3-week group), group B (5-week group) and group D (control group) by random number table.Group A received letrozole + CMC-Na mixture by gavage in the first 3 weeks and CMC-Na solution by gavage in the last 2 weeks, group B received letrozole + CMC-Na mixture by gavage for 5 weeks, and group D received CMC-Na solution by gavage for 5 weeks, and all three groups of rats were fed with normal diet.At the end of gavage, the body weight of rats in each group was observed, the histological changes of ovaries were observed by hematoxylin-eosin (HE) staining, the serum levels of estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), total cholesterol (TC), triglyceride (TG), fasting blood glucose (Glu), fasting insulin (FINS) and lipopolysaccharide (LPS) were measured by enzyme-linked immunosorbent assay (ELISA), and the LH/FSH ratio and insulin resistance index (HOMA IR) were calculated; the intestinal bacteria of rats were detected by 16S rRNA technique. Result: 1. Comparison of ovary histomorphology: Under light microscope, multiple luteum and oocytes were observed in mature follicles in group D, and granulosa cells were orderly arranged and multilayered, without cystic dilated follicles. There were no mature follicles in the ovarian tissues of group A and GROUP B. The follicles were irregular in structure and more cystic dilated follicles were visible. The number of granular cells in some follicles decreased or even disappeared. 2. Comparison of sex hormone levels: compared with group D, T level in group B was significantly increased (P < 0.001), and T level in group A had an upward trend (P > 0.05); The LH/FSH levels in group A and B were significantly increased (P < 0.001; P < 0.001). Compared with group A, E2 in group B was significantly decreased (P < 0.05) and T was significantly increased (P < 0.01). 3. Comparison of glucose and lipid metabolism levels: Compared with group D, TC levels in groups A and B were significantly increased (P < 0.01; P < 0.01). Compared with group A, TG in group B was significantly increased (P < 0.05). There were no significant differences in Glu, FINS and HOMA-IR levels among all groups. 4. Comparison of LPS levels: Compared with group D, the serum LPS levels of rats in groups A and B were significantly increased (P < 0.001; P < 0.01). 5. Intestinal flora analysis and comparison: At the phylum level, compared with group D, the abundance of Firmicutes in group B increased (P < 0.01), Firmicutes in group A showed an upward trend (P > 0.05), and the abundance of Bacteroidetes in groups A and B decreased (P < 0.05). At the genus level, compared with group D, Lactobacillus in group B increased (P < 0.01). The results of LEfSe analysis showed that there were differences in the composition of various intestinal bacteria among the three groups (LDA > 3).Conclusion: The phenotype of PCOS rats was related to the length of modeling, and the phenotypic characteristics of PCOS in rats at 5 weeks of modeling were more typical than those in rats at 3 weeks of modeling; PCOS can cause changes in intestinal flora, and the changes in the structure of intestinal flora between groups are related to different modeling duration.