目的:探讨芪芍抗纤方联合恩替卡韦对慢性乙型肝炎肝硬化患者肝功能、肝纤维化指标及TGF-β1(转化生长因子β1)、TIMP-1(基质金属蛋白酶抑制因子-1)的影响.方法:90 例慢性乙型肝炎肝硬化患者随机分2 组,对照组45 例口服恩替卡韦分散片,观察组45 例加服中药芪芍抗纤方,服药 3 个月.治疗前和治疗 3 个月后检测肝功能指标(ALT、AST、TBil)、肝纤四项(HA、PCⅢ、LN、Ⅳ-C)及血清TGF-β、TIMP-1 水平.结果:治疗3 个月后,2 组患者血清ALT、AST、TBil、HA、PCⅢ、LN、Ⅳ-C及TGF-β、TIMP-1 均显著下降(P＜0.05 或P＜0.01),观察组ALT、AST、TBil、HA、PCⅢ、LN、Ⅳ-C及TGF-β、TIMP-1 下降较对照组显著(均P＜0.05).结论:芪芍抗纤方联合恩替卡韦可改善乙肝肝硬化患的肝功能,减轻肝纤维化,降低TGF-β1 和TIMP水平.

Background: To clarify the inhibitory effect of Ercao Qinggan decoction (EQD) on acute liver failure (ALF) and its related mechanisms. Methods: HL-7702 hepatocytes were pretreated with TLR4 inhibitor CLI-095, glycogen synthase kinase 3 β (GSK3 β) inhibitor LiCl and different doses of EQD for 2 hours, and lipopolysaccharide (LPS) (10 μg/mL) for 24 hours. Cell apoptosis, TNF-α and IL-6 and GSK3β were detected by flow cytometry, immunofluorescence, quantitative polymerase chain reaction. After mice were gavaged with different concentrations of EQD for 12 days, ALF mouse models were established intraperitoneal injection of D-Gal/LPS. After 24 hours, the mice were euthanized and the liver tissue was stained with hematoxylin and eosin. Liver cell apoptosis, the serum levels of aspartate aminotransferase, alanine aminotransferase, TNF-α and IL-β were detected by terminal transferase-mediated dUTP nick end-labelling, enzyme linked immunosorbent assay, quantitative polymerase chain reaction, and Western blotting, respectively. These methods were also used to test the mRNA expression of Bax, Bcl-2 and the protein expression of GSK3β, p-Akt/Akt in livers. Results: CLI-095, LiCl, and EQD significantly inhibited apoptosis induced by LPS, the mRNA expression of IL-6, TNF-α and the nuclear translocation of GSK3β in HL-7702 hepatocytes. EQD dose-dependently inhibited hepatocyte apoptosis, the serum concentration of aspartate aminotransferase and ALT, the expression of TNF-α and IL-β, the ratio of p-GSK3β/GSK3β, p-Akt/Akt in alanine aminotransferase mice. Conclusion: EQD can inhibit hepatocyte apoptosis in ALF mice through regulating TLR4/PI3K/Akt/GSK3β signaling pathway.

目的:研究二草清肝汤(EQD)对内毒素脂多糖(LPS)联合D-氨基半乳糖(D-GalN)致小鼠急性肝损伤模型的:Toll样受体4(TLR4)信号通路的作用.方法:将21只BALB/C小鼠随机分为四个组:空白对照组(n=3)、内毒素模型组(n=7),均给予氯化钠溶液灌胃(3ml/kg);二草清肝汤小剂量组(n=6)、大剂量组(n=5)分别予生药含量为3.3g/ml、6.6g/ml的EQD溶液灌胃(3ml/kg).每日1次,连续12天.最后一天灌胃4h后,除空白对照组外,其余组所有小鼠均腹腔注射LPS和D-GalN混合液造模,腹腔注射24h后,处死所有小鼠,剖取其肝脏称重、HE染色显微观察病理;各组小鼠TLR4、NF-κB表达情况,应用实时荧光定量PCR(RT-qPCR)法从RNA水平检测、WB法从蛋白表达水平定量测定;TLR4蛋白表达应用免疫组化法.结果:内毒素性肝损伤小鼠肝重均较空白对照组低,加入二草清肝汤后小鼠肝重上升;病理结果显示空白组小鼠的肝小叶轮廓界限分明、清晰且完整,肝窦散布有序、较宽;肝上皮细胞排列规整、呈多边形;细胞质染色均匀、细胞核清晰、未见明显肿胀变性,库普弗细胞分布均匀,未见周部大量聚集.内毒素模型组中巨噬细胞聚集,肝小叶结构混乱、排列无序;肝细胞脂肪沉积、水肿明显较空白组严重;而二草清肝汤组病变则较前改善.免疫组化结果说明内毒素处理后的模型组中TLR4蛋白表达显著上升,加入大剂量二草清肝汤后TLR4稍有下降(P<0.05);PCR结果显示内毒素处理后的模型组中TLR4、NF-κB mRNA表达下降,加入二草清肝汤后明显上升(P<0.05).结论:二草清肝汤能够抑制肝细胞内TLR4信号通路的激活及相关炎症分子活化,改善LPS导致的肝损伤,其机制可能是EQD影响TLR4通路关键分子的蛋白翻译过程.

目的 探讨慢性乙型病毒性肝炎(以下简称乙肝)合并腹腔感染病原菌特征及危险因素.方法 选取温州市中医院收治的116例慢性乙肝患者作为研究对象,将47例腹腔感染者纳入观察组,69例无腹腔感染者纳入对照组.采用x2检验、多因素logistic回归分析慢性乙肝合并腹腔感染病原菌特征及危险因素.结果 观察组中检出病原菌50株.其中,革兰氏阴性菌31株,占62.00％,以肺炎克雷伯菌、大肠埃希菌、铜绿假单胞菌为主;革兰氏阳性菌17株,占34.00％,以金黄色葡萄球菌、屎粪肠球菌为主;真菌2株,占4.00％.糖尿病(OR=3.387)、住院时间≥30d(OR=5.643)、低蛋白血症(OR=3.156)、抗菌药物使用(多种)(OR=6.581)、有侵入性操作(OR=8.178)是慢性乙肝合并腹腔感染的独立危险因素(P＜0.05).结论 慢性乙肝合并腹腔感染患者的致病菌以革兰氏阴性菌为主,糖尿病、住院时间≥30 d、低蛋白血症、使用多种抗菌药物、有侵入性操作是其独立危险因素.

Objective:To investigate the effects of miRNA-26a (miR-26a) on the target gene HMGA2 on the proliferation and migration of hepatoma cells and the underlying mechanism. Methods:Liver cancer tissue samples ( n = 30) and adjacent normal tissue samples ( n = 30) pathologically confirmed by Wenzhou Hospital of Traditional Chinese Medicine between September 2018 and September 2019 were collected. MiR-26a mimics, control mimics (miR-Control), high-mobility group A2 protein (HMGA2) siRNA or negative control siRNA (Control) were transfected into human hepatoma cell lines HepG2 or Huh-7 cells. The expression of miR-26a in hepatocellular carcinoma tissue was detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR). MTT assay and scratch test were performed to determine the ability of cell proliferation and migration. RT-qPCR and western blotting were performed to detect miR-26a and HMGA2 mRNA expression. The relationship between miR-26a and HMGA2 mRNA was analyzed using Bioinformatics and luciferase reporter gene assay. Results:RT-qPCR results showed that the expression level of miR-26a in hepatocellular carcinoma tissue was 0.11 ± 0.02, which was significantly lower than that in normal tissues (0.25 ± 0.03, t = 21.268, P < 0.05). The expression level of miR-26a in stage III + IV was 0.05 ± 0.01, which was significantly lower than that in stage I + II (0.09 ± 0.01, t = 15.491, P < 0.05). Cell experiment showed that in the miR-26a group, the proliferation ability of Huh-7 cells was (3.10 ± 0.30) and (4.10 ± 0.40), and the proliferation ability of HepG2 cells was (3.08 ± 0.31) and (4.11 ± 0.40), which was significantly lower than that in the control group [(3.90 ± 0.40), (5.50 ± 0.60), (3.92 ± 0.41), (5.49 ± 0.58), t = 8.764, 10.634, 11.148, 10.728, all P < 0.05]. In the miR-26a group, the migration ability was (0.50 ± 0.06), (0.65 ± 0.07), which was significantly lower than that in the control group [(1.00 ± 0.10), (0.96 ± 0.10), t = 23.483, 13.910, both P < 0.05]. Bioinformatics and in vitro experiments showed that HMGA2 was a direct target of miR-26a. Restoring the expression of HMGA2 in miR-26a mimics-transfected cells, compared with that in the miR-26a group [(0.24 ± 0.02), (0.31 ± 0.03);(0.45 ± 0.05)], could significantly reverse the inhibitory effect of miR-26a on tumor cell proliferation and migration [(0.31 ± 0.03), (0.40 ± 0.04);(0.93 ± 0.08), t = 10.634, 9.859, 27.868, all P < 0.05). Conclusion:miR-26a inhibits the proliferation and migration of hepatoma cells by directly targeting HMGA2. The abnormal decrease of miR-26a and the increase of HMGA2 may be the important factors that participate in the occurrence and development of liver cancer.

目的 探讨五苓散联合多烯磷脂酰胆碱对非酒精性脂肪肝病患者的治疗效果.方法 选择2017年5月-2018年5月医院进行治疗的非酒精性脂肪肝的患者86例,按照随机数字的方法分为对照组(43例)和观察组(43例),所有患者给予常规治疗,对照组在常规方案的基础上使用多烯磷脂酰胆碱胶囊,观察组在对照组基础上使用五苓散,对两组治疗前后的中医症候积分进行对比.对两组治疗前后的谷丙转氨酶与谷草转氨酶进行对比.对两组治疗前后的总胆固醇与甘油三酯进行对比.对两组的治疗效果进行对比.结果 经过治疗后,两组的中医症候积分,谷丙转氨酶,谷草转氨酶,总胆固醇,甘油三酯均降低,观察组的降低幅度大于对照组(t=13.715、8.321、4.865、9.654、5.857、6.993、3.815、3.701,P＜0.05),观察组的治疗效果优于对照组(z=0-3.120,P＜0.05).结论 使用五苓散联合多烯磷脂酰胆碱治疗非酒精性脂肪肝病患者,可以显著缓解患者症状,改善患者的肝功能指标与血脂指标,提升治疗效果.

目的:探讨二草清肝汤对内毒素性肝损害模型大鼠肝脏Kupffer细胞的活化及功能依赖TLR4信号传导通路的影响.方法:120只雄性Wistar大鼠,随机分成空白组、模型组、二草清肝汤大剂量(20 g/kg)和小剂量组(10 g/kg),每组30只,连续灌胃给药12 d.末次给药4h后,除空白组外所有大鼠腹腔注射D-GalN (800 mg/kg)/LPS(0.04 mg/kg)诱导内毒素性肝损害,造模后1、2、4、6、12、24h每组取5只大鼠,Western印迹法检测Kupffer细胞Toll样受体-4(TLR-4)、TIR结构域的衔接蛋白(TRIF)、IL-1相关蛋白激酶(IRAK)、核因子-κB(NF-κB)蛋白表达变化.结果:造模后1、2h,各组大鼠TLR-4、TRIF、IRAK、NF-κB蛋白表达无显著差异.造模后4、6、12、24h,与空白组比较,模型组大鼠肝脏Kupffer细胞TLR-4、TRIF、IRAK、NF-κB蛋白表达明显增多(P＜0.05);与模型组比较,二草清肝汤大、小剂量组大鼠肝脏Kupffer细胞TLR-4、TRIF、IRAK、NF-κB蛋白表达明显减少(P＜0.05);大、小剂量组同时间比较无显著差异.结论:二草清肝汤可能通过抑制TLR-4信号转导通路的活性发挥其抗炎效应.

OBJECTIVE:We investigated the occurrence of recurrent aphthous stomatitis (RAS) among college students and its potential influence by dietary habits. METHODS:Study of dietary habits and RAS among students in Beijing University of Chinese Medicine was carried by homemade questionnaire. Multivariate binary logistic regression analysis was used to identify RAS risk factors and explore their relations. RESULTS:Among 1011 investigated college students, family history (odds ratio (OR) 1.678, 95% confidence intervals (CI) 1.192 to 2.364, p < 0.05), bed late (OR 1.515, 95% CI 1.005 to 2.285, p < 0.05), frequent thirst (OR 1.842, 95% CI 1.393 to 2.435, p < 0.001), and frequent drinking carbonated beverages (OR 1.369, 95% CI 1.029 to 1.821, p < 0.05) were independent risk factors for RAS, but preference for nuts (OR 0.607, 95% CI 0.448 to 0.824, p < 0.001) was a protective factor. There was no statistical difference in fruit intake between RAS and non-RAS groups (χ2 = 5.249, p > 0.05). CONCLUSIONS:Among college students, frequent drinking carbonated beverages or frequent thirst will increase its possibility, whereas preference for nuts provides protection. In addition, fruit intake does not have a positive effect.