Epigallocatechin gallate ameliorates tetrahydrochloride-induced liver toxicity in rats via inhibition of TGFβ / p-ERK/p-Smad1/2 signaling,antioxidant, anti-inflammatory activity

Chronic liver disease is a worldwide health problem. Carbon tetra hydrochloride is an environmental toxin which is regarded as highly toxic and a potential human carcinogen. It can cause liver damage through the generation of metabolites and production of free radicals. Green tea contains catechins such as Epigallocatechin gallate which has been found to reduce the inflammation, oxidative stress, and fibrosis in experimental animal models. Hence, it represents a good source to prevent or ameliorate several chronic diseases. Silymarin is extracted from milk thistle seeds and has been found to be an effective agent to reduce the oxidative stress and free radical production and thereby exert protective effects in chronic liver conditions. The present study was planned to keep in view the above-mentioned facts. We included thirty rats in our study and divided them into five groups, each having six rats and the study continued for 8 weeks. Group I received normal saline; Group 2 received i.p. CCl4 injections; Group 3 received CCl4 i.p. injection and Epigallocatechin gallate (EGCG) oral gavage, Group 4 received CCl4 i.p. injection and silymarin by oral gavage; and Group 5 received CCl4 i.p. injection and combined EGCG + silymarin by oral gavage. The study found that in group 2, CCl4 induced significant elevation of ALT and MDA and reduced GSH thereby signifying increased oxidative stress. CCl4 also significantly increased inflammatory (TNFα, NFκB, IL1β, and TGFβ) as well as fibrotic markers (p-ERK and p-Smad1/2 protein expression). EGCG and silymarin significantly reversed the previously mentioned parameters either alone or in combination; however, the effect was more pronounced in case of EGCG. We conclude that EGCG and silymarin possess liver protective effects through their antioxidant, anti-inflammatory, and antifibrotic action.     

重症监护室患儿心脏停搏后早期出现急性肾损伤的危险因素及预后分析

目的探讨心脏停搏（cardiac arrest，CA）患儿发生急性肾损伤（acute kidney injury，AKI）的相关因素及预后的影响因素。方法回顾性收集2016年6月—2021年6月湖南省儿童医院儿童重症监护室发生CA患儿的病历资料。按CA恢复自发循环（return of spontaneous circulation，ROSC）48 h内是否发生AKI分为AKI组（n=50）和非AKI组（n=113），AKI组按ROSC 7 d时预后情况分为存活组（n=21）和死亡组（n=29）。采用多因素logistic回归分析CA患儿早期发生AKI的相关因素及预后影响因素。结果CA后AKI发生率为30.7%（50/163）。AKI组7 d及28 d病死率分别为58.0%（29/50）、78.0%（39/50），非AKI组为31.9%（36/113）、58.4%（66/113）。多因素logistic回归分析显示，心肺复苏时间长（OR=1.164，95%CI：1.088~1.246，P<0.001）、基线血清白蛋白低（OR=0.879，95%CI：0.806~0.958，P=0.003）、CA前应用肾上腺素（OR=2.791，95%CI：1.119~6.961，P=0.028）与CA后AKI发生密切相关；基线小儿危重病例评分低（OR=0.761，95%CI：0.612~0.945，P=0.014）、CA前应用肾上腺素（OR=7.018，95%CI：1.196~41.188，P=0.031）、CA前机械通气（OR=7.875，95%CI：1.358~45.672，P=0.021）与CA后AKI患儿死亡密切相关。结论CA后ROSC患儿应密切监测血清白蛋白，尤其是心肺复苏时间长、基线小儿危重病例评分低、CA前应用肾上腺素、CA前机械通气者应及早识别和干预，以降低AKI发生率和病死率。     

Normobaric hyperoxia plays a neuroprotective role after cerebral ischemia by maintaining the redox homeostasis and the level of connexin43 in astrocytes

IntroductionAcute cerebral ischemia is caused by an insufficient blood supply to brain tissue. Oxygen therapy, which is able to aid diffusion to reach the ischemic region, has been regarded as a possible treatment for cerebral ischemia. Recent animal and pilot clinical studies have reported that normobaric hyperoxia (NBO) showed neuroprotective effects if started soon after the onset of stroke. However, little is known about the role and mechanism of NBO treatment in astrocytes. Connexin43, one of the main gap junction proteins in astrocytes, is extremely sensitive to hypoxia and oxidative stress after cerebral ischemia.AimsIn the present study, we used sutures to develop an ischemia/reperfusion model in rats to mimic clinical recanalization and investigated the role of connexin43 in NBO‐treated stroke rats, as well as the underlying mechanism of NBO therapy.ResultsNormobaric hyperoxia treatment maintained the homeostasis of oxidoreductases: glutathione peroxidase 4 (GPX4) and NADPH oxidase 4 (two important oxidoreductases) and rescued the ischemia/reperfusion‐induced downregulation of connexin43 protein in astrocytes. Furthermore, NBO treatment attenuated cerebral ischemia‐induced cytochrome c release from mitochondria and was involved in neuroprotective effects by regulating the GPX4 and connexin43 pathway, using Ferrostatin‐1 (an activator of GPX4) or Gap27 (an inhibitor of connexin43).ConclusionsThis study showed the neuroprotective effects of NBO treatment by reducing oxidative stress and maintaining the level of connexin43 in astrocytes, which could be used for the clinical treatment of ischemic stroke.     

靶向β-catenin/TCF4相互作用小分子抑制剂荧光偏振高通量筛选模型的建立与应用

基于荧光偏振(fluorescence polarization, FP)原理,建立并应用以β-catenin/TCF4 (T-cell factor 4)相互作用为靶标的小分子抑制剂荧光偏振高通量筛选模型获得苗头化合物。利用大肠杆菌原核表达系统,原核表达和分离纯化重组人β-catenin,以酶联免疫吸附实验(enzyme-linked immunosorbent assay, ELISA)进行生物学活性鉴定。以异硫氰酸荧光素(fluorescence isothiocyanate, FITC)标记的TCF4多肽为荧光探针,通过优化FITC-TCF4与β-catenin反应浓度,建立并应用靶向β-catenin/TCF4相互作用小分子抑制剂荧光偏振高通量筛选模型进行苗头化合物筛选。利用大肠杆菌原核表达系统成功进行了重组人β-catenin原核表达与分离纯化。ELISA实验证实了纯化的重组人β-catenin具有良好的生物学活性。选用20 nmol·L-1FITC-TCF4和100 nmol·L-1β-catenin,成功建立了Z’因子为0.88的荧光偏振高通量筛选模型。应用本筛选模型进...     

S100A4蛋白在膀胱癌中的表达及其临床意义

目的探讨膀胱癌中S100A4的表达及意义。方法采用免疫组化S-P法检测65例膀胱癌中S100A4的表达情况,分析其表达与临床病理特征之间的关系。结果S100A4的表达与组织学分级、淋巴结转移呈正相关(P<0.05)。结论膀胱癌中S100A4的表达与肿瘤的进展密切相关,是判断膀胱癌生物学行为、预测转移趋势的有价值的指标。     

高效液相色谱法测定复方胆通胶囊中羟甲香豆素的含量

目的:用高效液相色谱法测定复方胆通胶囊中羟甲香豆素的含量.方法:高效液相色谱法,采用SWG(200 mm×4.6 mm,5 μm)柱,以萘为内标,甲醇-水(3:1)为流动相,流速为1.00 mL·min-1, 检测波长为252 nm.结果:羟甲香豆素在17～68 μg·mL-1浓度范围内呈线性关系(r=0.999 9,n=7).该法回收率99.4%,RSD为0.05%(n=5).结论:该方法简单、快速、准确.     

Association of plasma bone morphogenetic protein‐4 levels with arterial stiffness in hypertensive patients

BackgroundArterial stiffness interacts with hypertension, becoming an early marker of hypertension‐mediated target organ damage. This study aimed to assess the association between plasma concentrations of bone morphogenetic protein‐4 (BMP‐4) and arterial stiffness during hypertension.MethodsUsing cardio‐ankle vascular index (CAVI) to determine arterial stiffness status, 204 individuals with essential hypertension were classified into two groups, high CAVI (abnormal) group (n = 94) and low (normal) CAVI group (n = 110). Data were collected including clinical characteristics and laboratory measurements. Plasma levels of BMP‐4 were tested by using ELISA analysis.ResultsPlasma levels of BMP‐4 were substantially greater in high CAVI group than that in low CAVI group [38.51 (31.79–50.83) pg/mL vs. 31.15 (29.38–32.37) pg/mL; p < 0.001]. As shown by spearman correlation analysis, BMP‐4 concentrations were correlated with CAVI values in hypertensive individuals (r = 0.406, p < 0.001). After adjustment for potential confounders, elevated BMP‐4 levels were related with high CAVI (OR, 1.070; 95% CI, 1.003–1.108; p < 0.001). The best BMP‐4 cutoff value for identifying high CAVI, as determined by ROC curve analysis, was 33.34 pg/mL (AUC, 0.751; 95% CI, 0.683–0.818; p < 0.001).ConclusionPlasma levels of BMP‐4 are increased in hypertensive individuals with high CAVI. Elevated BMP‐4 levels are strongly correlated with higher CAVI values, implying a predictive value of BMP‐4 in arterial stiffness during hypertension.     

Richter转化患者的克隆同源性检测及其分子生物学特征分析

目的探索Richter转化（RT）患者的克隆同源性、临床与分子生物学特征。方法回顾性分析南京医科大学第一附属医院血液科（浦口慢淋中心）2019年1月至2021年12月确诊的18例RT患者慢性淋巴细胞白血病/小淋巴细胞淋巴瘤（CLL/SLL）及转化弥漫大B细胞淋巴瘤（DLBCL）的免疫球蛋白重链可变区（IGHV）基因片段使用及IGHV-D-J重排模式，鉴定患者克隆同源性。结合患者初诊及转化时的临床信息及分子检测，分析RT患者的高危因素。结果18例RT患者转化时中位年龄56.5（41～75）岁。其中17例转化为DLBCL，1例转化为霍奇金淋巴瘤（HL）。17例RT DLBCL患者中，15例（88.2％）患者DLBCL与CLL/SLL克隆同源；2例（11.8％）患者与CLL/SLL克隆非同源。其中11例患者转化前未接受治疗的CLL/SLL样本与转化后DLBCL样本配对的二代测序（NGS）结果显示突变频率最高的基因均为EGR2、TP53、NOTCH1；但部分患者转化时出现上述基因突变的新获得或丢失，提示存在克隆演变；10例布鲁顿酪氨酸激酶（BTK）抑制剂治疗后转化的患者中4例出现BTK突变。上述突变可能为促进转化的高危因素；此外，TP53、EGR2突变可能为包含新药联合方案治疗RT的不良预后因素。结论本中心转化DLBCL患者大多为克隆同源性转化；推荐有条件的中心开展相关检测。转化前未治CLL/SLL与转化后DLBCL组织的突变谱有一定异质性。     

166例双表达弥漫大B细胞淋巴瘤临床特征与预后分析

目的探讨双表达弥漫大B细胞淋巴瘤（DLBCL）的临床特征与预后。方法回顾性分析2016年1月至2020年12月北京大学第三医院收治的166例双表达DLBCL患者的临床资料，分析患者的临床特征、生存和预后因素。结果共收集DLBCL患者410例，其中双表达淋巴瘤（DEL）166例（40.5％）。男82例，女84例，中位诊断年龄63.5（21～95）岁。110例（66.3％）患者初诊年龄≥60岁，106例（106/163，65.0％）患者诊断时LDH升高，74例（74/160，46.2％）诊断时β₂-微球蛋白（β₂-MG）≥3 mg/L，107例（107/163，65.6％）结外受累数目≥2个，65例（65/166，39.2％）有B症状，131例（131/165，79.4％）初诊时分期为Ⅲ、Ⅳ期，41例（41/161，25.5％）初诊时国际预后指数（IPI）评分0～2分，38例（38/161，23.6％）初诊时IPI评分3分，82例（82/161，50.9％）初诊IPI评分4～5分。DEL患者中9例（9/56，16.1％）具有MYD88和CD79B突变。单因素分析显示，年龄≥60岁（P＝0.004）、β₂-MG水平升高（P＝0.002）、IPI评分高（P＝0.003）与较差的总生存（OS）相关，β₂-MG水平升高（P＝0.031）、LDH水平升高（P＝0.017）、分期Ⅲ～Ⅳ期（P＝0.001）、IPI评分高（P＝0.013）、免疫组化p53突变型（P＝0.049）和PIM1突变（P＝0.039）与较差的无进展生存（PFS）相关。多因素分析显示，IPI评分4～5分（HR＝2.622，95％CI 1.398～4.917，P＝0.003）是影响DEL患者OS的独立危险因素。生存分析显示，DEL与非DEL患者的PFS率差异有统计学意义（65.6％对75.1％，P＝0.002），而OS率的差异无统计学意义（81.8％对83.6％，P＝0.226）。在DEL患者中，REPOCH方案的总有效率高于RCHOP或RCHOP样方案（81.5％对63.4％，P＝0.004）。结论DEL是一组侵袭性较强的淋巴瘤，具有较差的PFS，采用REPOCH方案化疗可能会改善患者的整体预后。     

舒洛地特通过抑制NOX4/NLRP3通路减轻高糖诱导的视网膜血管损伤

目的:观察舒洛地特(SDX)在糖尿病状态下对视网膜微血管内皮细胞的保护作用。方法:(1)采用高脂喂养及腹腔注射链脲佐菌素的方法构建C57BL/6小鼠的2型糖尿病模型,对小鼠腹腔注射SDX或等体积生理盐水治疗12周。用伊文思蓝法检测视网膜微血管渗漏及微血管密度改变;用Western blot、免疫荧光等方法检测各组小鼠视网膜中NLRP3炎症小体成分、紧密连接蛋白1(ZO-1)及NADPH氧化酶4(NOX4)的表达情况。(2)用高糖处理人视网膜微血管内皮细胞(HRMECs),并与梯度浓度的SDX共处理;用腺病毒感染HRMECs以过表达NOX4或者用siRNA转染HRMECs以敲减NOX4,观察NLRP3炎症小体成分、NOX4、ZO-1和VE-钙黏着蛋白(VE-Cad)的表达情况,以及活性氧簇(ROS)的生成情况。结果:(1)SDX处理增加糖尿病小鼠视网膜中ZO-1蛋白的表达,抑制糖尿病小鼠视网膜微血管渗漏,增加糖尿病小鼠视网膜微血管密度和节细胞数量,并抑制NLRP3炎症小体的活化(P<0. 05)。(2)在高糖刺激的HRMECs中,SDX处理增加ZO-1和VE-Cad蛋白的表达,显著...