Pathophysiological relationship between hypoxia associated oxidative stress,Epithelial-mesenchymal transition,stemness acquisition and alteration of Shh/ Gli-1 axis during oral sub-mucous fibrosis and oral squamous cell carcinoma

Oral sub-mucous fibrosis (OSF) is a pathophysiological state of oral cavity or oropharynx having a high chance of conversion to oral squamous cell carcinoma (OSCC). It involves fibrotic transformation of sub-epithelial matrix along with epithelial abnormalities. The present work aims to unveil the mechanistic domain regarding OSF to OSCC conversion exploring the scenario of hypoxia associated oxidative stress, epithelial-mesenchymal transition (EMT), metastasis and stemness acquisition. The study involves histopathological analysis of the diseased condition along with the exploration of oxidative stress status, assessment of mitochondrial condition, immunohistochemical analysis of HIF-1α, E-cadherin, vimentin, ERK, ALDH-1, CD133, Shh, Gli-1 and survivin expressions in the oral epithelial region together with the quantitative approach towards collagen deposition in the sub-epithelial matrix. Oxidative stress was found to be associated with type-II EMT in case of OSF attributing the development of sub-epithelial fibrosis and type-III EMT in case of OSCC favoring malignancy associated metastasis. Moreover, the acquisition of stemness during OSCC can also be correlated with EMT. Alteration of Shh and Gli-1 expression pattern revealed the mechanistic association of hypoxia with the phenotypic plasticity and disease manifestation in case of OSF as well as OSCC. Shh/ Gli-1 signaling can also be correlated with survivin mediated cytoprotective phenomenon under oxidative stress. Overall, the study established the correlative network of hypoxia associated oxidative stress, EMT and manifestation of oral pre-cancerous and cancerous condition in a holistic approach that may throw rays of hope in the therapeutic domain of the concerned diseases.     

Efficient screening of predictive biomarkers for individual treatment selection

The development of molecular diagnostic tools to achieve individualized medicine requires identifying predictive biomarkers associated with subgroups of individuals who might receive beneficial or harmful effects from different available treatments. However, due to the large number of candidate biomarkers in the large‐scale genetic and molecular studies, and complex relationships among clinical outcome, biomarkers, and treatments, the ordinary statistical tests for the interactions between treatments and covariates have difficulties from their limited statistical powers. In this paper, we propose an efficient method for detecting predictive biomarkers. We employ weighted loss functions of Chen et al. to directly estimate individual treatment scores and propose synthetic posterior inference for effect sizes of biomarkers. We develop an empirical Bayes approach, namely, we estimate unknown hyperparameters in the prior distribution based on data. We then provide efficient screening methods for the candidate biomarkers via optimal discovery procedure with adequate control of false discovery rate. The proposed method is demonstrated in simulation studies and an application to a breast cancer clinical study in which the proposed method was shown to detect the much larger numbers of significant biomarkers than existing standard methods.     

牛磺酸上调基因1(TUG1)对肝纤维化的作用及其机制*

目的:探讨牛磺酸上调基因1(TUG1)在肝纤维化中的作用机制。方法:按照文献建立TGF-β1(5 ng/ml)刺激的活化肝星状细胞模型和经典的1%DMN(1 ml/kg/d)致大鼠肝纤维化模型,将肝纤维化大鼠和活化肝星状细胞(HSC)均分为模型对照组、阴性对照组(沉默TUG1阴性对照)、siRNA干扰组(TUG1基因沉默组)。实验结束后利用苏木精-伊红(HE)染色检测大鼠肝脏组织病理变化;采用逆转录-聚合酶链反应(RT-PCR)法、蛋白免疫印记(Western blot)分别测定大鼠肝组织及活化肝星状细胞中α-平滑肌肌动蛋白(α-SMA)、TUG1、I型胶原蛋白(collagenI)、基质金属蛋白酶-2(MMP-2)、金属蛋白酶组织抑制因子(TIMP-1)、Smad2、Smad3表达水平。结果:肝组织病理学检查显示,沉默TUG1能够明显缓解肝脏纤维化病理改变,Western blot结果显示,沉默TUG1能够显著降低大鼠肝组织和活化肝星状细胞中TUG1、α-SMA、collagenI、MMP-2、TIMP-1、Smad2、Smad3基因与蛋白表达水平(P＜0.05)。与模型对照组相比,阴性对照组的TUG1、α-SMA的蛋白与基因水平明显升高(P＜0.05)。与模型对照组和阴性对照组相比,siRNA干扰组中TUG1, α-SMA, collagenI, MMP-2, TIMP-1, Smad2 and Smad3的蛋白和基因水平显著降低(P＜0.05),而在模型对照组和阴性对照组中TUG1, α-SMA, collagenI, MMP-2, TIMP-1, Smad2 and Smad3的蛋白和基因表达水平之间差异无显著性。结论:TUG1在肝纤维化组织和活化的肝星状细胞中显著上调,沉默TUG1可能通过抑制转化生长因子-β1(TGF-β1)/Smad信号通路改善1%DMN致大鼠肝纤维化病理损伤,降低活化肝星状细胞中纤维化相关蛋白水平,发挥抗肝纤维化的作用。     

百草枯通过SOCE途径促进肺纤维化的发生

摘要 目的：探讨SOCE对百草枯（PQ）致肺纤维化过程的具体调控作用。方法：体外培养肺泡上皮A549细胞,设置分组为对照组、PQ组和PQ+SKF96365组。对照组不作任何处理;PQ组给予800 μM的PQ溶液处理细胞24 h;PQ+SKF96365组先加入10 μM的SKF96365预处理细胞2 h,然后再加800 μM PQ处理24 h。免疫荧光检测PQ染毒后SOCE相关蛋白的活化和胞内分布、NFATc1的表达和入核情况。清洁级小鼠40只,随机分为空白对照组、PQ组、SKF96365组和PQ+SKF96365组。PQ组及PQ+SKF96365组给予百草枯（20 mg/kg）一次性腹腔注射,对照组给予等量生理盐水;SKFS96365组及PQ+SKF96365组每天腹腔注射一次10 mg/kg的SKF96365溶液,持续3天,行HE染色、Masson染色观察肺组织病理状态改变以及胶原纤维的变化情况。结果：与正常组相比,PQ染毒组STIM1蛋白活化出现寡聚化现象并且ORAI1、TRPC1膜定位显著;PQ引起细胞质内NFATc1信号向细胞核内转移,密度差异具有统计学意义（P<0.01）,使用SOCE抑制剂后NFATc1信号减弱向细胞质内转移（P<0.01）。与对照组相比,PQ中毒的小鼠肺泡结构被破坏,肺间质大量胶原纤维沉积;与PQ组相比,PQ+SKF96365组肺泡结构相对完整并且间质胶原纤维沉积减少。结论：PQ中毒可活化SOCE并通过增加NFAT入核激活下游转录信号,促进肺纤维化的发生。     

微血管减压术联合感觉根部分切断术对原发性三叉神经痛患者疼痛评分、生活质量及睡眠状况的影响

目的:探讨微血管减压术(MVD)联合感觉根部分切断术(PSR)对原发性三叉神经痛(TN)患者疼痛评分、生活质量及睡眠状况的影响。方法:回顾性分析2015年2月~2019年3月期间我院收治的80例原发性TN患者的临床资料,根据手术方式的不同将患者分为对照组(n=40,MVD治疗)和研究组(n=40,MVD联合PSR治疗),比较两组患者疼痛评分、生活质量、围术期指标、睡眠状况、并发症发生情况以及复发率。结果:两组患者治疗后视觉疼痛模拟量表(VAS)评分均较治疗前下降,且研究组低于对照组(P<0.05)。两组患者治疗后生活质量量表(SF-36)各维度评分均较治疗前升高,且研究组高于对照组(P<0.05)。两组患者治疗后匹兹堡睡眠质量指数表(PSQI)各项目评分均较治疗前升高,且研究组高于对照组(P<0.05)。研究组住院时间短于对照组,手术时间长于对照组(P<0.05);两组术中出血量比较无统计学差异(P>0.05)。研究组的并发症总发生率低于对照组(P<0.05)。两组随访期间复发率比较差异无统计学意义(P>0.05)。结论:MVD联合PSR治疗原发性TN,虽然手术时间较长,但是在减轻患者疼痛、改善患者生活质量及睡眠状况等方面效果显著,能够降低并发症发生率。     

慢性乙肝病毒感染患者肝纤维化、肝功能及Th1/Th2细胞因子水平变化及恩替卡韦干预研究

摘要 目的：研究慢性乙肝病毒（HBV）感染患者肝纤维化、肝功能及Th1/Th2细胞因子水平的变化情况,并分析恩替卡韦干预对慢性HBV感染患者相关指标的影响。方法：选择2016年4月至2018年1月我院收治的120例慢性HBV感染者为研究对象,其中慢性乙肝轻度50例、中度24例、重度21例、肝硬化者25例,并选择同期于我院进行体检的50例健康者为健康对照组,分析不同病情慢性HBV感染患者血清中的肝纤维化指标[透明质酸（HA）、Ⅳ型胶原（ⅣC）,层粘连蛋白（LN）、III型前胶原肽（PC III）]、肝功能指标[丙氨酸转氨酶（ALT）、天冬氨酸转移酶（AST）]及Th1/Th2细胞因子[干扰素-γ（IFN-γ）、白介素-2（IL-2）、白介素-4（IL-4）、白介素-6（IL-6）]的变化,慢性HBV感染患者按照随机数字表法分为观察组和对照组,对照组给予还原型谷胱甘肽和复方甘草酸苷等常规护肝治疗,观察组在对照组基础上另口服恩替卡韦分散片治疗,对比两组治疗前后各指标水平。结果：慢性HBV感染患者的肝纤维化指标、肝功能指标及Th1/Th2细胞因子IL-4、IL-6等指标水平均显著高于健康对照组,且随着病情的加重呈逐渐上升趋势;而IFN-γ和IL-2水平均低于健康对照组,随着病情的加重而逐渐降低（P<0.05）。治疗后两组血清肝纤维化指标（HA、ⅣC、LN、PC III）、肝功能指标（ALT、AST）以及IL-4、IL-6水平均显著低于治疗前,且观察组低于对照组（P<0.05）。治疗后两组血清IFN-γ、IL-2水平均高于治疗前,且观察组高于对照组（P<0.05）。结论：慢性HBV感染患者存在肝纤维化水平上升、肝功能受损及Th1/Th2细胞失衡,恩替卡韦联合常规护肝治疗可有效纠正患者的Th1/Th2细胞失衡,减轻患者的肝纤维化,改善其肝功能。     

Using vaginal discharge score (VDS) grading system to evaluate the effect of clinical endometritis on reproductive performance of dairy cows in China

Clinical endometritis (CE) is a major cause in affecting the reproductive performance of dairy cows. The objectives of this study were to ascertain the prevalence of CE and to evaluate the effect of CE on reproductive performance in dairy cows using vaginal discharge score (VDS) grading system. 803 dairy cows were examined by vaginoscope with 4-point VDS at 26 ± 3 days in milk (DIM) and classified into six groups: non-endometritis with VDS 0 (control; CON), endometritis with VDS 1 (MEM), non-treated endometritis with VDS 2 (NTME), treated endometritis with VDS 2 (TME), non-treated endometritis with VDS 3 (NTPE), and treated endometritis with VDS 3 (TPE). Cows in TME and TPE groups were treated with 200 mL of 50% dextrose solution by intrauterine infusion. The prevalence of CE was 33% at 26 ± 3 DIM. Binary logistic regression analysis revealed cows in MEM, NTME and NTPE groups had a less likelihood of first artificial insemination (AI) pregnancy than those in CON group (P < 0.05). Kaplan-Meier survival curves for days open were statistically different (P = 0.004). In Cox regression model, cows in NTME and NTPE groups had a reduced pregnancy rate than those in CON group (P < 0.05). The hazard of pregnancy in NTME group was lower than that in TME group (P = 0.044). Similarly, it was lower for the hazard of pregnancy in NTPE group than in TPE group (P = 0.048). Cows in MEM, NTME, and NTPE groups required more services per pregnancy than those in CON group (P < 0.05). In conclusion, CE examined by the VDS grading system impaired reproductive performance, and mild endometritis with VDS 1 should be treated in the early postpartum period to ameliorate fertility in dairy herds.     

Inhibition of cyclooxygenase-2 enhanced intestinal epithelial homeostasis via suppressing β-catenin signalling pathway in experimental liver fibrosis

The intestinal barrier dysfunction is crucial for the development of liver fibrosis but can be disturbed by intestinal chronic inflammation characterized with cyclooxygenase-2 (COX-2) expression. This study focused on the unknown mechanism by which COX-2 regulates intestinal epithelial homeostasis in liver fibrosis. The animal models of liver fibrosis induced with TAA were established in rats and in intestinal epithelial–specific COX-2 knockout mice. The impacts of COX-2 on intestinal epithelial homeostasis via suppressing β-catenin signalling pathway were verified pharmacologically and genetically in vivo. A similar assumption was tested in Ls174T cells with goblet cell phenotype in vitro. Firstly, disruption of intestinal epithelial homeostasis in cirrhotic rats was ameliorated by celecoxib, a selective COX-2 inhibitor. Then, β-catenin signalling pathway in cirrhotic rats was associated with the activation of COX-2. Furthermore, intestinal epithelial–specific COX-2 knockout could suppress β-catenin signalling pathway and restore the disruption of ileal epithelial homeostasis in cirrhotic mice. Moreover, the effect of COX-2/PGE2 was dependent on the β-catenin signalling pathway in Ls174T cells. Therefore, inhibition of COX-2 may enhance intestinal epithelial homeostasis via suppression of the β-catenin signalling pathway in liver fibrosis.     

Macrophage migration inhibitory factor regulates joint capsule fibrosis by promoting TGF-β1 production in fibroblasts

Joint capsule fibrosis caused by excessive inflammation results in post-traumatic joint contracture (PTJC). Transforming growth factor (TGF)-β1 plays a key role in PTJC by regulating fibroblast functions, however, cytokine-induced TGF-β1 expression in specific cell types remains poorly characterized. Macrophage migration inhibitory factor (MIF) is a proinflammatory cytokine involved in inflammation- and fibrosis-associated pathophysiology. In this study, we investigated whether MIF can facilitate TGF-β1 production from fibroblasts and regulate joint capsule fibrosis following PTJC. Our data demonstrated that MIF and TGF-β1 significantly increased in fibroblasts of injured rat posterior joint capsules. Treatment the lesion sites with MIF inhibitor 4-Iodo-6-phenylpyrimidine (4-IPP) reduced TGF-β1 production and relieved joint capsule inflammation and fibrosis. In vitro, MIF facilitated TGF-β1 expression in primary joint capsule fibroblasts by activating mitogen-activated protein kinase (MAPK) (P38, ERK) signaling through coupling with membrane surface receptor CD74, which in turn affected fibroblast functions and promoted MIF production. Our results reveal a novel function of trauma-induced MIF in the occurrence and development of joint capsule fibrosis. Further investigation of the underlying mechanism may provide potential therapeutic targets for PTJC.     

Development and validation of glycolysis-related prognostic score for prediction of prognosis and chemosensitivity of pancreatic ductal adenocarcinoma

Pancreatic ductal adenocarcinoma (PDAC) is a lethal malignancy with aggressive biological behaviour. Its rapid proliferation and tumour growth require reprogramming of glucose metabolism or the Warburg effect. However, the association between glycolysis-related genes with clinical features and prognosis of PDAC is still unknown. Here, we used the meta-analysis to correlate the hazard ratios (HR) of 106 glycolysis genes from MSigDB by the cox proportional hazards regression analysis in 6 clinical data sets of PDAC patients to form a training cohort, and a single group of PDAC patients from the TCGA, ICGC, Arrayexpress and GEO databases to form the validation cohort. Then, a glycolysis-related prognosis (GRP) score based on 29 glycolysis prognostic genes was established in 757 PDAC patients from the training composite cohort and validated in 267 ICGC-CA validation cohort (all P < .05). In addition, including PADC, the prognostic value was also confirmed in other 7 out of 30 pan-cancer cohorts. The GRP score was significantly related to specific metabolism pathways, immune genes and immune cells in the patients with PADC (all P < .05). Finally, by combining with immune cells, the GRP score also well-predicted the chemosensitivity of patients with PADC in the TCGA cohort (AUC = 0.709). In conclusion, this study developed a GRP score for patients with PDAC in predicting prognosis and chemosensitivity for PDAC.