鲎血细胞蛋白营养学评价及其水解肽体外活性

对鲎血细胞蛋白(TTBCP)进行营养学评价,并测定TTBCP水解物的体外抗氧化性、α-葡萄糖苷酶和乙酰胆碱酯酶的抑制活性。结果显示:TTBCP中必需氨基酸优于成人的FAO/WHO推荐模式,基本符合小孩的FAO/WHO推荐模式,其中必需氨基酸与总氨基酸的比值(E/T)为31.2%,氨基酸评分(AAS)为68.1,蛋白质效率比(PER)小于2.0。TTBCP水解肽对α-葡萄糖苷酶的半数抑制浓度(IC_(50))为0.199 mg/mL,抑制率最高达到83.8%,对乙酰胆碱酯酶同样有抑制作用;TTBCP水解肽能清除超氧阴离子自由基、DPPH自由基和羟自由基,当浓度小于0.75 mg/mL的范围时,还原能力大于维生素C。由此可见,TTBCP具有复合功效,可作为糖尿病的保健食品和药品开发利用。

全血细胞计数和高密度脂蛋白胆固醇比值对心力衰竭的预测价值

目的 探讨中性粒细胞与高密度脂蛋白胆固醇(HDL-C)比值(NHR)、单核细胞/HDL-C比值(MHR)和淋巴细胞与HDL-C比值(LHR)在心力衰竭(HF)的预测价值。方法 根据心功能分级将慢性HF患者420例分为美国纽约心脏病学会(NYNA)心功能分级Ⅲ级组230例和Ⅳ级组190例。根据有无急性肾损伤分为肾损伤组160例和无肾损伤组260例。根据出院后6个月有无主要心血管不良事件(MACE)分为预后不良组50例和预后良好组370例。分别于入院后24、72 h,检测血常规、血脂、肾功能、N末端B型脑钠尿肽前体(NT-proBNP)、可溶性肿瘤因子2抑制剂(sST2)水平;计算NHR、MHR、LHR和估算肾小球滤过率(eGFR)。于入院后24 h、出院时,测定左心室射血分数(LVEF)、6 min步行距离(6MWD)、明尼芬达生活质量量表(MLHFQ),出院后6个月,随访MACE。结果 入院后24 h, NYNAⅣ级组、肾损伤组、预后不良组NHR、MHR、LHR、sST2、NT-proBNP、MLHFQ评分分别高于NYNAⅢ级组、无肾损伤组、预后良好组,LVEF、6MWD分别低于NYNAⅢ级组、无肾损伤组、预后良好组,差异有统计学意义(P<0.05);尿素氮(BUN)、血肌酐(Scr)、eGFR两组间差异无统计学意义(P>0.05);入院后72 h, NYNAⅣ级组、肾损伤组、预后不良组NHR、MHR、LHR、sST2、NT-proBNP、MLHFQ评分、BUN、Scr分别高于NYNAⅢ级组、无肾损伤组、预后良好组,LVEF、6MWD、eGFR分别低于NYNAⅢ级、无肾损伤组、预后良好组,差异有统计学意义(P<0.05);入院后24 h,各组NHR、MHR、LHR、sST2、NT-proBNP高于入院后72 h, MLHFQ评分高于出院时;各组LVEF、6MWD低于出院时,差异有统计学意义(P<0.05)。相关性分析:入院后24 h患者NHR、MHR、LHR与NT-proBNP呈正相关(r=0.894、0.847、0.931,均P=0.001);与入院后72 h eGFR呈负相关(r=-0.868、-0.731、-0.806,P=0.001、0.001、0.007)。Logistic回归分析显示,入院后24 h NHR、MHR、LHR升高患者发生急性肾损伤的危险性增加。受试者工作特征(ROC)曲线结果显示,NHR、MHR、LHR联合检测曲线下面积为0.821,对HF发生MACE预测价值最大。结论 全血细胞计数和HDL-C比值升高,HF患者发生急性肾损伤危险性增加,对HF短期预后有预测价值。

HAX-1调控人宫颈癌Hela的细胞增殖

目的探讨造血细胞特异性蛋白1(HS-1)相关蛋白X-1(HAX-1)对宫颈癌Hela细胞增殖的调控作用。方法以人宫颈癌Hela细胞系为研究模型,采用脂质体介导法将HAX-1 siRNAs和阴性对照siRNA转染Hela细胞并构建HAX-1沉默细胞系(HAX-1沉默组),以HAX-1高表达质粒和pcDNA3.1空载质粒转染Hela细胞并构建HAX-1过表达细胞系(HAX-1过表达组)。采用实时荧光定量PCR(RT-qPCR)和Western blot检测转染后Hela细胞中HAX-1的表达情况;EdU试验检测转染后的细胞增殖能力;MTT试验检测转染后的细胞活力;流式细胞术检测HAX-1对转染后Hela细胞周期和细胞凋亡的变化;RT-qPCR检测转染后Ki-67、c-Myc、BCL-2/BAX基因的表达水平。结果沉默HAX-1后,Hela细胞中HAX-1 mRNA和蛋白质的水平显著降低,而过表达HAX-1后,Hela细胞中HAX-1 mRNA和蛋白质的水平显著升高,差异均有统计学意义(P<0.05)。过表达HAX-1后,HAX-1过表达组中EdU阳性细胞数目下降,细胞活力降低,G1期细胞比例增加,S期细胞比例降低,细胞凋亡率上升,增殖相关基因Ki67表达降低,BCL-2/BAX基因显著降低(P<0.05),而HAX-1沉默组则具有相反的表现。结论HAX-1的表达抑制了Hela细胞增殖,并通过BCL-2/BAX途径诱导其凋亡。

血细胞比容与白蛋白差值评估急性百草枯中毒程度的临床研究

目的探讨血细胞比容与白蛋白差值(HCT-ALB)评估急性百草枯(PQ)中毒患者中毒程度的临床价值。方法选择云南省第二人民医院2018年1月至2019年12月收治的急性PQ中毒患者作为研究对象,以同期健康体检者作为对照。收集患者的一般情况、中毒量、中毒时间,以及入院时未输注血液制品、未静脉输液、未行血液净化时测量的HCT和血清ALB,计算HCT-ALB差值。根据入院时尿PQ快速半定量检测结果将患者分成PQ低浓度组(0~10 mg/L)和PQ高浓度组(30~100 mg/L)。分析中毒时间、中毒量以及HCT-ALB差值与急性PQ中毒程度的关系,并采用Spearman法进行等级相关性分析。结果共纳入295例急性PQ中毒患者,其中PQ低浓度组118例,PQ高浓度组177例;以及性别、年龄与PQ患者相匹配的200例健康体检者(健康对照组)。PQ低浓度组中毒时间长于PQ高浓度组〔h:11.0(6.0,60.0)比8.0(5.0,20.5),P<0.01〕,但中毒量显著低于PQ高浓度组〔mL:10.0(5.8,15.0)比40.0(20.0,80.0),P<0.01〕。PQ低浓度组和PQ高浓度组HCT、HCT-ALB差值均显著高于健康对照组〔HCT:(43.14±4.41)%、(43.54±5.40)%比(42.14±2.15)%,HCT-ALB差值:3.59±6.26、5.94±7.80比-7.26±3.55,均P<0.01〕,而ALB则显著低于健康对照组(g/L:39.54±5.74、37.60±7.15比49.40±3.41,均P<0.01)。随着尿液PQ浓度升高,HCT、HCT-ALB差值进一步升高,ALB进一步下降,PQ高浓度组与PQ低浓度组组间比较差异均有统计学意义〔HCT:(43.54±5.40)%比(43.14±4.41)%,HCT-ALB差值:5.94±7.80比3.59±6.26,ALB(g/L):37.60±7.15比39.54±5.74,均P<0.05〕。急性PQ中毒患者中毒程度与中毒时间、ALB呈负相关(r值分别为-0.195、-0.695,均P<0.01),而与中毒量、HCT、HCT-ALB差值呈正相关(r值分别为0.650、0.256、0.737,均P<0.01),其中HCT-ALB差值与中毒程度的相关性最强。结论HCT-ALB差值可反映急性PQ中毒患者的中毒程度,间接揭示急性PQ中毒患者体内微血管的病理改变。

低表达人血细胞特异蛋白1相关蛋白X1对人结肠癌细胞增殖的影响

目的观察低表达人血细胞特异蛋白1相关蛋白X1(HAX1)对结肠癌细胞增殖的影响。方法利用HAX1特异性低表达小干扰RNA(siHAX1)瞬时转染结肠癌细胞SW48,通过Western blot验证低表达成功,采用细胞计数试剂盒-8(CCK-8)、克隆形成实验观察HAX1对SW48细胞的影响,利用Western blot检测细胞周期蛋白(Cyclin) D1表达。结果Western blot证实在结肠癌细胞中低表达HAX1模型的成功建立,CCK-8实验结果表明,对照组(siCon)48 h结肠癌细胞吸光度(A)值(1.264±0.020)较各实验组(siHAX1)A值(1.122±0.057、1.021±0.033、0.989±0.049)差异有统计学意义(P<0.05),即实验组增殖能力降低。72 h时,对照组A值(1.332±0.019),各实验组A值(1.199±0.040、1.081±0.022、1.068±0.026),组间比较差异有统计学意义(P<0.05),说明72 h时实验组增殖能力仍降低。克隆形成实验显示,对照组(siCon)克隆形成数目[(713±40)个]明显多于各实验组克隆形成数目[(422±33)、(558±37)、(505±23)个],且差异有统计学意义(P<0.05),说明实验组克隆形成能力降低。Western blot证实在细胞低表达HAX1蛋白后,可明显降低Cyclin D1蛋白的表达。结论在结肠癌细胞中,低表达HAX1可通过抑制Cyclin D1的表达抑制肿瘤细胞增殖。

外周血细胞角质蛋白-19可作为鼻咽癌转移的标志

鼻咽癌生长迅速且容易转移。台北Yand-Ming大学的研究者认为,外周血中细胞角质蛋白-19的mRNA可以作为鼻咽癌患者肿瘤转移的指标。 研究对象为57名未经治疗、无转移灶的进展型鼻咽癌患者。研究者以反转录多聚酶链反应分析法(RT-PCR)

新生大鼠缺血缺氧性脑损伤组织中HAX-1、Caspase-3、bcl-2表达变化及意义

目的观察新生大鼠缺氧缺血性脑损伤(HIBD)组织中血细胞特异蛋白1相关蛋白X-1(HAX-1)、Caspase-3、bcl-2的表达,并探讨其意义。方法 128只大鼠随机分为观察组和对照组各64只,观察组采用Rice法制备HIBD模型,对照组仅分离左侧颈总动脉不结扎。分别于造模后0、2、6、12、24、48、72 h断头取脑,采用Western blot法检测大脑皮层HAX-1、Caspase-3、bcl-2蛋白。结果与对照组同时点比较,观察组HAX-1于造模后6、12、18、48 h表达升高,Caspase-3于造模后6、12、18 h表达升高,bcl-2于造模后2、6、12 h表达升高,P均<0.05;观察组造模后12、18 h的HAX-1的表达水平均高于造模后6 h,P均<0.05。结论新生大鼠HIBD模型大脑皮层中存在HAX-1、Caspase-3、bcl-2高表达,三者共同参与了HIBD的发生、发展。

HAX-1和MMP-7在大肠癌中的表达及临床意义

目的:研究造血细胞特异蛋白1相关蛋白X1(haematopoietic lineage cell specific protein 1-associated protein X-1, HAX-1)和基质金属蛋白酶-7(matrix metalloproteinase-7, MMP-7)在大肠癌中的表达情况及与临床病理特征和预后的关系。方法:应用免疫组化法检测99例经手术切除的大肠癌标本和38例正常肠黏膜组织中HAX-1和MMP-7的表达,观察HAX-1和MMP-7在大肠癌中的表达情况及与临床病理特征和预后的关系,分析两者在大肠癌组织中表达的相关性。结果:大肠癌组织中HAX-1和MMP-7的阳性表达率分别为81.82%和53.54%,与正常黏膜组织比较差异有统计学意义(P<0.05)。HAX-1表达水平与肿瘤分期、淋巴结转移和肿瘤分化程度相关(P<0.05),MMP-7与肿瘤分期、淋巴结转移相关(P<0.05)。HAX-1和MMP-7在大肠癌组织中的表达呈正相关。HAX-1及MMP-7的表达与大肠癌患者的预后密切相关。结论:HAX-1及MMP-7在大肠癌中均高表达,在大肠癌的发病过程中它们的高表达具有协同效应,与大肠癌淋巴结转移、临床分期及预后密切相关,并可作为判断大肠癌患者预后的重要指标之一。

Regulation of hematopoiesis and the hematopoietic stem cell niche by Wnt signaling pathways

Hematopoietic stem cells （HSCs） are a rare population of cells that are responsible for life-long generation of blood cells of all lineages. In order to maintain their numbers, HSCs must establish a balance between the opposing cell fates of self-renewal （in which the ability to function as HSCs is retained） and initiation of hematopoietic differentiation. Multiple signaling pathways have been implicated in the regulation of HSC cell fate. One such set of pathways are those activated by the Wnt family of ligands. Wnt signaling pathways play a crucial role during embryogenesis and deregulation of these pathways has been implicated in the formation of solid tumors. Wnt signaling also plays a role in the regulation of stem cells from multiple tissues, such as embryonic, epidermal, and intestinal stem cells. However, the function of Wnt signaling in HSC biology is still controversial. In this review, we will discuss the basic characteristics of the adult HSC and its regulatory microenvironment, the ＂niche＂, focusing on the regulation of the HSC and its niche by the Wnt signaling pathways.

HAX-1通过调控MDM2/SMAD3通路对TGF-β1诱导的特发性肺间质纤维化的作用机制

目的探讨造血细胞特异性蛋白1相关蛋白X-1(HAX-1)对转化生长因子-β1(TGF-β1)诱导的特发性肺间质纤维化的作用及机制。方法将A549细胞分为对照组(无处理)、TGF-β1组(用5μg·L^(-1)的TGF-β1诱导48 h)、NC si组(用阴性对照siRNA转染细胞,再用5μg·L^(-1)的TGF-β1诱导48 h)、HAX-1 si组(用HAX-1 siRNA转染细胞,再用5μg·L^(-1)的TGF-β1诱导48 h)、HAX-1 si+pcDNA3.1组(用HAX-1 siRNA转染细胞,再用pcDNA3.1空质粒转染细胞,再用5μg·L^(-1)的TGF-β1诱导48 h)、HAX-1 si+MDM2组(用HAX-1 siRNA转染细胞,再用MDM2过表达质粒pcDNA3.1-MDM2转染细胞,再用5μg·L^(-1)的TGF-β1诱导48 h)、HAX-1 si+MDM2+DMSO组(用HAX-1 siRNA转染细胞,再用MDM2过表达质粒pcDNA3.1-MDM2转染细胞,向细胞培养液中加入溶剂DMSO处理细胞30 min,再用5μg·L^(-1)的TGF-β1诱导48 h)、HAX-1 si+MDM2+SIS3组(用HAX-1 siRNA转染细胞,再用MDM2过表达质粒pcDNA3.1-MDM2转染细胞,用终浓度为1μmol·L^(-1)溶于DMSO的SIS3处理细胞30 min,再用5μg·L^(-1)的TGF-β1诱导48 h)。相应处理结束后,采用倒置相差显微镜观察细胞形态变化;Western blot检测HAX-1、鼠双微体基因2(MDM2)、SMAD3、p-SMAD3、E-钙黏蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)和Ⅰ型胶原α1链(COL1A1)蛋白表达水平。结果TGF-β1诱导细胞后,细胞由鹅卵石形或多角形转变为梭形、纺锤形,E-cadherin蛋白表达降低(P<0.05),HAX-1、α-SMA和COL1A1蛋白表达升高(P<0.05)。干扰HAX-1可提高E-cadherin蛋白表达(P<0.05),抑制MDM2、p-SMAD3、α-SMA和COL1A1蛋白表达(P<0.05),并且细胞形态又转变为鹅卵石形或多角形。MDM2过表达抵消HAX-1干扰对p-SMAD3、E-cadherin、α-SMA和COL1A1蛋白表达的影响(P<0.05)。SMAD3抑制剂SIS3处理后细胞形态又转变为鹅卵石形或多角形,E-cadherin蛋白表达升高(P<0.05),α-SMA和COL1A1蛋白表达下降(P<0.05)。结论HAX-1在TGF-β1诱导的A549细胞中高表达,HAX-1敲低可通过MDM2/SMAD3通路抑制TGF-β1诱导的肺泡细胞纤维化过程。

Wnt5a participates in hepatic stellate cell activation observed by gene expression profile and functional assays

AIM:To identify differentially expressed genes in quiescent and activated hepatic stellate cells(HSCs)and explore their functions.METHODS:HSCs were isolated from the normal Sprague Dawley rats by in suit perfusion of collagenase and pronase and density Nycodenz gradient centrifugation.Total RNA and mRNA of quiescent HSCs,and cultureactivated HSCs were extracted,quantified and reversely transcripted into cDNA.The global gene expression profile was analyzed by microarray with Affymetrix rat genechip.Differentially expressed genes were annotated with Gene Ontology(GO)and analyzed with Kyoto encyclopedia of genes and genomes(KEGG)pathway using the Database for Annotation,Visualization and Integrated Discovery.Microarray data were validated by quantitative real-time polymerase chain reaction(qRTPCR).The function of Wnt5a on human HSCs line LX-2 was assessed with lentivirus-mediated Wnt5a RNAi.The expression of Wnt5a in fibrotic liver of a carbon tetrachloride(CCl4)-induced fibrosis rat model was also analyzed with Western blotting.RESULTS:Of the 28 700 genes represented on this chip,2566 genes displayed at least a 2-fold increase or decrease in expression at a P<0.01 level with a false discovery rate.Of these,1396 genes were upregulated,while 1170 genes were downregulated in culture-activated HSCs.These differentially expressed transcripts were grouped into 545 GO based on biological process GO terms.The most enriched GO terms included response to wounding,wound healing,regulation of cell growth,vasculature development and actin cytoskeleton organization.KEGG pathway analysis revealed that Wnt5a signaling pathway participated in the activation of HSCs.Wnt5a was significantly increased in cultureactivated HSCs as compared with quiescent HSCs.qRTPCR validated the microarray data.Lentivirus-mediated suppression of Wnt5a expression in activated LX-2 resulted in significantly impaired proliferation,downregulated expressions of typeⅠcollagen and transforming growth factor-β1.Wnt5a was upregulated in the fibrotic liver of a CCl4-induced fibrosis rat model.CONCLUSION:Wnt5a is involved in the activation of HSCs,and it may serve as a novel therapeutic target in the treatment of liver fibrosis.

Uptake of albumin nanoparticle surface modified with glycyrrhizin by primary cultured rat hepatocytes

AIM: To investigate the uptake difference between bovine serum albumin nanoparticle (BSA-NP) and bovine serum albumin nanoparticles with their surface modified byglycyrrhizin (BSA-NP-GL) and to develop a novel hepatocyte targeting BSA-NP-GL based on active targeting technology mediated by specific binding site of GL on rat cellular membrane. METHODS: Calcein loaded bovine serum albumin nanoparticles (Cal-BSA-NP) were prepared by desolvation process. Glycyrrhizin was conjugated to the surface reactive amino groups (SRAG) of Cal-BSA-NP by sodium periodate oxidization, which resulted in calcein-loaded bovine serum albumin nanoparticles with their surface modified by glycyrrhizin (Cal-BSA-NP-GL). The morphology of the two types of prepared nanoparticles (NP) was observed by transmission electron microscopy. The diameter of NP was measured with a laser particle size analyzer. The interaction between Cal-BSA-NP-GL and primary cultured hepatocytes was studied through cellular uptake experiments. The uptake amount of Cal-BSA-NPGL and Cal-BSA-NP by rat hepatocytes was determinedby fluorospectrophotometry. Uptake characteristics were investigated through experiments of competitive inhibition of specific binding site of GL. RESULTS: Both Cal-BSA-NP-GL and Cal-BSA-NP had regular spherical surfaces. The average diameter of CalBSA-NP-GL and Cal-BSA-NP was 77 and 79 nm respectively. The uptake amount of the two NP by hepatocytes reached its maximum at 2 h after incubation. The uptake amount of Cal-BSA-NP-GL by rat hepatocytes was 4.43-fold higher than that of Cal-BSA-NP. There was a significant difference in the uptake of Cal-BSA-NP-GL and Cal-BSA-NP by hepatocytes (P＜0.01). The uptake of Cal-BSA-NP-GL was inhibited when GL was added previously to isolated rat hepatocytes, and the uptake of Cal-BSA-NP was not affected by GL.CONCLUSION: A binding site of GL is present on the surface of rat hepatocytes, BSA-NP-GL may be internalized via this site by hepatocytes and can be used as a drug carrier for active targeting of delivery drugs to hepatocytes.

Pancreatitis-associated protein:From a lectin to an anti-inflammatory cytokine

Pancreatitis-associated protein (PAP) was discovered in the pancreatic juice of rats with acute pancreatitis. PAP is a 16 kDa secretory protein structurally related to the C-type lectins although classical lectin-related function has not been reported yet. Then, it was demonstrated that PAP expression may be activated in some tissues in a constitutive or injury- and inflammation-induced manner. More recently, it has been found that PAP acts as an anti-inflammatory factor in vitro and in vivo. PAP expression can be induced by several pro- and anti-inflammatory cytokines and by itself through a JAK/STAT3-dependent pathway. PAP is able to activate the expression of the anti-inflammatory factor SOCS3 through the JAK/STAT3-dependent pathway. The JAK/STAT3/SOCS3 pathway seems to be a common point between PAP and several cytokines. Therefore, it is reasonable to propose that PAP is a new anti- inflammatory cytokine.

益气化瘀汤联合^(131)I治疗老年分化型甲状腺癌的疗效及对免疫功能、CK19、MUC1表达的影响

目的探讨益气化瘀汤联合^(131)I治疗老年分化型甲状腺癌(DTC)的效果及对免疫功能、血细胞角蛋白19(CK19)、多态性上皮黏蛋白(MUC1)表达的影响。方法选取2016年4月—2019年4月该院收治的老年DTC患者300例为研究对象,采用随机数字表法将其分为对照组和观察组各150例,对照组采用^(131)I治疗;观察组在对照组的基础上采用益气化瘀汤治疗。比较两组临床疗效、治疗前和治疗2个月后免疫功能、CK19和MUC1水平、甲状腺功能指标[游离三碘甲状腺原氨酸(FT3)、血清游离甲状腺素(FT4)、促甲状腺激素(TSH)、甘油三酯(TG)],并进行1年随访,记录两组复发率。结果观察组总有效率高于对照组[88.67%(133/150)vs 75.33%(113/150)](P<0.05);治疗后2个月后,两组CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)、FT3、FT4水平均升高,CD8^(+)、TSH、TG水平及CK19和MUC1阳性表达率均降低(P<0.05);且观察组CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)、FT3、FT4水平高于对照组,CD8^(+)、TSH、TG水平及CK19和MUC1阳性表达率低于对照组(P<0.05);观察组1年复发率低于对照组[1.33%(2/150)vs 13.33%(20/150)](P<0.05)。结论益气化瘀汤联合^(131)I治疗老年DTC的效果显著,可降低CK19、MUC1水平,提高免疫功能和甲状腺功能,降低复发率。

Effect of serum concentration on adhesion of monocytic THP-1 cells onto cultured EC monolayer and EC-SMC co-culture

Background:The adhesion of monocytes to the endothelium following accumulation of low-density lipoprotein (LDL) in subendothelial spaces is an important step in the development of intimal hyperplasia in arterially implanted vein grafts and atherosclerosis in both animals and humans. However, it is not well known how serum factors affect the adhesion of monocytes. Methods: We have studied the effect of fetal calf serum (FCS), which we considered a source of LDL, on the adhesion of monocytes to endothelial cells (ECs) by using human monocytic THP-1 cells and both a monolayer of cultured bovine aortic endothelial cells (EC monoculture) and a co-culture with bovine aortic smooth muscle cells (EC-SMC co-culture). Results: It was found that the addition of FCS to the medium greatly affected the adhesion of THP-1 cells, and the higher the concentration of FCS in the medium, the greater the adhesion of THP-1 cells to endothelial cells. Adhesion of THP-1 cells to an EC-SMC co-culture was approximately twofold greater than that to an EC monoculture, and after adhering to endothelial cells, many THP-1 cells trans-migrated into the layer of smooth muscle cells. Conclusion: The results suggest that the elevation of the LDL (cholesterol) level in blood provides a favorable condition for the development of intimal hyperplasia and atherosclerosis by promoting the adhesion of monocytes to the endothelium and their subsequent migration into subendothelial spaces.

Effects of ischemic preconditioning on cyclinD1 expression during early ischemic reperfusion in rats

AIM： To observe the effect of ischemic preconditioning on cyclinD1 expression in rat liver cells during early ischemic reperfusion.METHODS： Fifty-four SD rats were randomly divided into ischemic preconditioning group （IP）, ischemia/ reperfusion group （IR） and sham operation group （SO）. The IP and IR groups were further divided into four sub-groups （n = 6）. Sham operation group （SO） served as the control group （n = 6）. A model of partial liver ischemia/reperfusion was used, in which rats were subjected to liver ischemia for 60 min prior to reperfusion. The animals in the IP group underwent ischemic preconditioning twice for 5 min each time prior to the ischemia/reperfusion challenge. Alter 0, 1, 2, and 4 h of reperfusion, serum and liver tissue in each group were collected to detect the level of serum ALT, liver histopathology and expression of cyclinD1 mRNA and protein. Flow cytometry was used to detect cell cycle as the quantity indicator of cell regeneration. RESULTS： Compared with IR group, IP group showed a significantly lower ALT level in 1h to 4h sub-groups （P 〈 0.05）. Proliferation index（PI） indicated by the S-phase and G2/M-phase ratio [（S＋G2/M）/（G0/G1＋S＋G2/M）] was significantly increased in IP group at 0 and 1 h （26.44 ± 7.60% vs 18.56 ± 6.40%,41.87 ± 7.27% vs 20.25 ± 6.70%, P 〈 0.05）. Meanwhile, cyclinD1 protein expression could be detected in IP group. But in IR group, cyclinD1 protein expression occurred 2 h alter reperfusion. The expression of cyclinD1 mRNA increased significantly in IP group at 0 and 1h （0.568 ± 0.112 vs 0.274 ± 0.069, 0.762 ± 0.164 vs 0.348 ± 0.093,P 〈 0.05）.CONCLUSION： Ischemic preconditioning can protect liver cells against ischemia/reperfusion injury, which may be related to cell proliferation and expression of cyclinD1 during early ischemic reperfusion.

Crosstalk between angiogenesis, cytokeratin-18, and insulin resistance in the progression of non-alcoholic steatohepatitis

AIM: To elucidate the possible crosstalk between angiogenesis, cytokeratin-18 (CK-18), and insulin resistance (IR) especially in patients with non-alcoholic steatohepatitis (NASH).METHODS: Twenty-eight patients with NASH and 11 with simple fatty liver disease (FL) were enrolled in this study and underwent clinicopathological examination. The measures of angiogenesis, CK-18, and IR employed were CD34-immunopositive vessels, CK-18immunopositive cells, and homeostasis model assessment of IR (HOMA-IR), respectively. The correlations of these factors with NASH were elucidated.RESULTS: Significant development of hepatic neovascularization was observed only in NASH, whereas almost no neovascularization could be observed in FL and healthy liver. The degree of angiogenesis was almost parallel to liver fibrosis development, and both parameters were positively correlated. Similarly, CK-18expression and HOMA-R were signifi cantly increased in NASH as compared with FL and healthy liver. Furthermore, CK-18 and HOMA-IR were also positively correlated with the degree of neovascularization. CONCLUSION: These results indicate that the crosstalk between angiogenesis, CK-18, and IR may play an important role in the onset and progression of NASH.

New insights into the role of PML in tumour suppression

The PML gene is involved in the t（15;17） translocation of acute promyelocytic leukaemia （APL）, which generates the oncogenic fusion protein PML （promyelocytic leukaemia protein）-retinoic acid receptor alpha. The PML protein localises to a subnuclear structure called the PML nuclear domain （PML-ND）, of which PML is the essential structural component. In APL, PML-NDs are disrupted, thus implicating these structures in the pathogenesis of this leukaemia. Unexpectedly, recent studies indicate that PML and the PML-ND play a tumour suppressive role in several different types of human neoplasms in addition to APL. Because of PML＇s extreme versatility and involvement in multiple cellular pathways, understanding the mechanisms underlying its function, and therefore role in tumour suppression, has been a challenging task. In this review, we attempt to critically appraise the more recent advances in this field and propose new avenues of investigation.

益气化瘀汤联合131Ⅰ治疗老年分化型甲状腺癌的疗效及预后相关因素分析

目的探讨益气化瘀汤联合131Ⅰ治疗老年分化型甲状腺癌(differentiated thyroid cancer,DTC)的疗效及预后相关因素。方法选取2017年3月—2019年4月该院收治的老年DTC患者328例,采用随机数法将其分为对照组(164例)和观察组(164例)。对照组采用基础治疗+131Ⅰ进行治疗,观察组在对照组的基础上采用益气化瘀汤进行治疗,两组均连续治疗2个月。比较两组治疗2个月后的患者的临床疗效;对比两组治疗前、治疗2个月后的血细胞角蛋白19(CK19)、多态性上皮黏蛋白(MUC1)表达水平及甲状腺功能相关指标水平;并分析影响患者复发的因素。结果治疗2个月后,观察组总有效率为87.20%(143/164),显著高于对照组76.83%(126/164),(P <0.05)。与治疗前比较,治疗2个月后,两组CK19、MUC1阳性表达率及甲状腺激素(TSH)、甲状腺球蛋白(TG)水平均降低,且观察组均显著低于对照组(P <0.05);两组游离三碘甲状腺原氨酸(FT3)、游离甲状腺素(FT4)水平均升高,且观察组均显著高于对照组(P <0.05)。多因素Logistic回归分析结果显示,年龄≥65岁、性别为男性、分期为Ⅲ、Ⅳ期、肿瘤直径≥4 cm、TG水平≥0.3ng/mL以及未使用131Ⅰ为老年分化型甲状腺癌患者复发的危险因素(P <0.05)。结论益气化瘀汤联合131Ⅰ治疗DTC可明显降低患者CK19、MUC1阳性表达率,同时可改善患者甲状腺功能,且分析疾病复发的因素可预防或减少患者疾病复发。

Science Letters:Serum protein fingerprinting coupled with artificial neural network distinguishes glioma from healthy population or brain benign tumor

To screen and evaluate protein biomarkers for the detection of gliomas (Astrocytoma grade Ⅰ-Ⅳ) from healthy individuals and gliomas from brain benign tumors by using surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) coupled with an artificial neural network (ANN) algorithm. SELDI-TOF-MS protein fingerprinting of serum from 105 brain tumor patients and healthy individuals, included 28 patients with glioma (Astrocytoma Ⅰ-Ⅳ), 37 patients with brain benign tumor, and 40 age-matched healthy individuals. Two thirds of the total samples of every compared pair as training set were used to set up discriminating patterns, and one third of total samples of every compared pair as test set were used to cross-validate; simultaneously, discriminate-cluster analysis derived SPSS 10.0 software was used to compare Astrocytoma grade Ⅰ-Ⅱ with grade Ⅲ-Ⅳ ones. An accuracy of 95.7%, sensitivity of 88.9%, specificity of 100%, positive predictive value of 90% and negative predictive value of 100% were obtained in a blinded test set comparing gliomas patients with healthy individuals; an accuracy of 86.4%, sensitivity of 88.9%, specificity of 84.6%, positive predictive value of 90% and negative predictive value of 85.7% were obtained when patient's gliomas was compared with benign brain tumor. Total accuracy of 85.7%, accuracy of grade Ⅰ-Ⅱ Astrocytoma was 86.7%, accuracy ofⅢ-Ⅳ Astrocytoma was 84.6% were obtained when grade Ⅰ-Ⅱ Astrocytoma was compared with grade Ⅲ-Ⅳ ones (discriminant analysis). SELDI-TOF-MS combined with bioinformatics tools, could greatly facilitate the discovery of better biomarkers. The high sensitivity and specificity achieved by the use of selected biomarkers showed great potential application for the discrimination of gliomas patients from healthy individuals and glioma from brain benign tumors.