PHLPP2在人胶质瘤中的表达及意义

目的研究PHLPP2在人正常脑组织以及脑胶质瘤组织中的表达及其临床意义。方法选取青岛大学附属医院与安丘人民医院神经外科自2014年12月至2016年10月间手术切除并经病理证实的人脑胶质瘤标本56例,其中I级7例,Ⅱ级19例,Ⅲ级21例,胶质母细胞瘤9例。另取20例因脑创伤行内减压术患者的正常脑组织标本作为对照。应用RT-PCR、Western-blotting分别检测各标本中PHLPP2m RNA和PHLPP2蛋白的表达。结果 RT-PCR和Western-blotting检测结果显示正常脑组织中PHLPP2 mRNA和PHLPP2蛋白表达最高,不同级别胶质瘤组织PHLPP2 mRNA和PHLPP2蛋白表达差异有统计学意义(P<0.05),且随着肿瘤病理级别的增高,胶质瘤组织中PHLPP2 mRNA和PHLPP2蛋白表达值降低,差异有统计学意义(P<0.05)。结论 PHLPP2在正常脑组织和不同级别的脑胶质瘤组织中都有表达,其表达值与肿瘤的病理分级呈负相关,PHLPP2可能作为一个肿瘤抑制基因在胶质瘤的发生、发展中起到关键性的作用。     

CD123在急性白血病骨髓干祖细胞中的表达及其临床意义

目的检测细胞表面抗原CD123（IL-3受体α链）在急性白血病（AL）患者骨髓干祖细胞中的表达,并探讨其与患者预后的关系。方法采用流式细胞术检测43例初治的AL患者及13例正常对照组的骨髓CD34阳性细胞中CDl23的表达情况;同时根据AL患者的年龄、细胞遗传学改变、发病时外周血白细胞数量将其按不同预后分为高危、标危、低危3组,其中高危15例,标危15例,低危13例,分析CD123表达与AL患者预后的相关性。结果43例AL患者CD34＋CDl23＋/CD34＋平均水平为38.24%,显著高于对照组平均表达水平1.90%;3组AL患者CD34＋CDl23＋/CD34＋均显著高于对照组（P〈0.05）;高危组患者CD34＋CDl23＋/CD34＋显著高于标危组和低危组,差异具有统计学意义（P〈0.05）;标危组患者CD34＋CDl23＋/CD34＋显著高于低危组,差异具有统计学意义（P〈0.05）。结论在AL患者骨髓干祖细胞中,CD123异常过度表达;在高危组、标危组、低危组的表达逐渐降低,提示CD123的表达与AL患者的预后相关,临床可通过检测该指标判断AL患者预后。     

骨髓增生异常综合征Th1/Th2亚群测定意义分析

目的 分析Th1/Th2亚群测定对骨髓增生异常综合征(Myelodysplastic syndrome,MDS)分型及预后的价值.方法 搜集我院确诊的MDS患者60例,分为SLD/MLD组和EB-1/EB-2组,按照IPSS-R标准分为极低危组,低危组,中危组,高危组,极高危组.选取25例健康人作为对照组.流式细胞仪测定外周血PE-IFN-γ+(Th1)、PE-IL-4+(Th2)水平,统计学分析结果在各组间的表达,与CDCD45RA、CD45RO做相关性分析.结果 SLD/MLD组Th1水平、Th1/Th2水平高于健康对照组(P<0.01),EB-1/EB-2组Th1水平、Th1/Th2水平低于健康对照组(P<0.01),Th2水平高于健康对照组(P<0.01);IPSS-R危险度分组:低危组(P<0.01)、中危组(P<0.01)Th1、Th1/Th2高于健康对照组,高危组(P<0.01)极高危组(P<0.05)Th1、Th1/Th2低于健康对照组,Th2高于健康对照组.Th1与CD45RO呈正相关关系(r=0.514,P<0.01).结论 MDS不同阶段Th1/Th2亚群极化方向不同,SLD/MLD组向Th1极化,细胞免疫亢进,EB-1/EB-2组向Th2极化,体液免疫增强;随着IPSS-R危险度积分的升高,细胞免疫在中高危组之间由亢进转为被抑制,免疫监视逃逸,肿瘤细胞积累,引起疾病进展.     

骨髓增生异常综合征Th1/Th2亚群测定意义分析

目的 分析Th1/Th2亚群测定对骨髓增生异常综合征(Myelodysplastic syndrome,MDS)分型及预后的价值.方法 搜集我院确诊的MDS患者60例,分为SLD/MLD组和EB-1/EB-2组,按照IPSS-R标准分为极低危组,低危组,中危组,高危组,极高危组.选取25例健康人作为对照组.流式细胞仪测定外周血PE-IFN-γ+(Th1)、PE-IL-4+(Th2)水平,统计学分析结果在各组间的表达,与CDCD45RA、CD45RO做相关性分析.结果 SLD/MLD组Th1水平、Th1/Th2水平高于健康对照组(P<0.01),EB-1/EB-2组Th1水平、Th1/Th2水平低于健康对照组(P<0.01),Th2水平高于健康对照组(P<0.01);IPSS-R危险度分组:低危组(P<0.01)、中危组(P<0.01)Th1、Th1/Th2高于健康对照组,高危组(P<0.01)极高危组(P<0.05)Th1、Th1/Th2低于健康对照组,Th2高于健康对照组.Th1与CD45RO呈正相关关系(r=0.514,P<0.01).结论 MDS不同阶段Th1/Th2亚群极化方向不同,SLD/MLD组向Th1极化,细胞免疫亢进,EB-1/EB-2组向Th2极化,体液免疫增强;随着IPSS-R危险度积分的升高,细胞免疫在中高危组之间由亢进转为被抑制,免疫监视逃逸,肿瘤细胞积累,引起疾病进展.     

运动平板试验Duke评分对胸痛患者危险分层及预后的价值

目的评价运动平板试验Duke评分(DTS)对胸痛患者危险分层及预后评估的应用价值。方法选择169例运动试验阳性和可疑阳性同时行冠状动脉造影的患者为研究对象,按Duke评分分为DTS低危组(Duke≥5分,n=35)、中高危组(DTS<+5,n=134),比较两组患者冠状动脉病变和临床特点,随访两年比较2组发生主要不良心血管事件(MACE)的概率,分析DTS预测MACE的价值。结果在中高危组中限制性心绞痛发作例数、ST段偏移≥1 mm例数、运动时ST段改变涉及导联数目、ST段偏移值均明显高于DTS低危组(P<0.05);而运动持续时间和运动最大心率明显低于DTS低危组(P<0.05)。DTS与冠脉病变的严重程度相关,冠脉造影结果显示低危组冠脉造影Gensini积分低于中高危组(P<0.0001);胸痛患者有随访资料的169例,失访9例,160例完成随访,失访率为5.33%。MACE主要发生在随访1个月,在DTS中高危组中发生MACE明显高于DTS低危组(P<0.05);logistic多因素回归分析显示,DTS是心血管事件发生的危险因素(OR=1.434,1.278¹.609;Waldχ2=37.7076,P<0.0001)。结论运动试验DTS与冠状动脉病变狭窄程度高度相关;DTS中高危组发生MACE明显高于低值组,应用该评分可以有效地对临床中胸痛患者进行危险分层及预后的评估,对于DTS中高危组的患者应迅速进行评估并考虑早期行再灌注治疗。     

血清N末端B型钠尿肽原水平对原发性高血压病患者危险分层相关性研究

目的:通过原发性高血压病(EH)患者不同危险分层血清N末端B型钠尿肽原(NT-proBNP)水平的变化,探讨其相关性及临床意义。方法:用放射免疫法测定了155例原发性高血压病患者和36例非高血压老年患者的NT-proBNP水平,并进行对照统计分析。结果显示:EH组血清N-BNP水平显著高于对照组(P<0.01),男女组间无统计学差异。低危组、中危组、高危组、很高危险组间均递增呈显著正相关(r=0.873,P<0.01),但低危组与对照组无显著差异(P>0.05)。伴心脑肾并发症组指标均显著高于无并发症组(P<0.01)。结论:EH患者存在明显的血清NT-proBNP水平的变化,且这种变化与EH危险分组明显相关,提示NTproBNP的变化与病情进展、预后相关,对高血压病分层评估及预后有指导作用,可能成为高血压病危险分层新指标。     

LOX-1对急性冠脉综合征的早期预测价值

目的:探讨血凝素样氧化低密度脂蛋白受体-1(LOX-1)对急性冠脉综合征的早期预测价值.方法:选择行冠脉动脉造影的患者75例,根据临床情况分为正常对照组(NC组)22例、不稳定型心绞痛组(UA组)28例和急性心肌梗死组(AMI组)25例,根据SYNTAX积分分为高危组(19例)和低危组(34例).采用酶联免疫法测定患者血清LOX-1水平.结果:AMI组LOX-1水平高于UA组和NC组,UA组LOX-1水平高于NC组,差异均有统计学意义(P＜0.05或P＜0.01).高危组患者LOX-1水平(107.43±35.18)ng/L高于低危组(86.90±31.27) ng/L,差异有统计学意义(t=2.19,P＜0.05).结论:LOX-1是预测急性冠脉综合征很有潜力的生物学标志物.     

儿童急性淋巴细胞白血病中期因子表达的临床研究

目的:检测儿童急性淋巴细胞白血病(ALL)患者中期因子(MK)的表达,探讨MK在儿童ALL发病中的意义.方法:应用实时定量PCR(real-time PCR),对124例进展期和缓解期ALL患者、15例正常儿童骨髓标本进行MK mRNA的检测.根据患者初治时外周血白细胞数、年龄、免疫分型及皮质激素预治疗反应等因素把进展期患者分为标危组、中危组和高危组.结果:进展期、缓解期及正常对照组之间MK水平比较差异有统计学意义(P < 0.01).MK mRNA在B-ALL亚型中表达显著高于T-ALL亚型及正常对照(均P < 0.01),而T-ALL与正常对照之间MK mRNA的表达差异无统计学意义(P > 0.05).在ALL危险度各分层中,标危组和中危组的MK表达水平明显高于高危组(P < 0.01 或P < 0.05),标危组与中危组之间差异无统计学意义(P = 0.32).MK表达水平与患儿年龄、性别、血浆乳酸脱氢酶等未显示出相关性(均P > 0.05).高白细胞组(WBC ≥ 25×10~9/L)MK的表达较白细胞不增高组(WBC < 25×10~9/L)明显降低(P < 0.05).结论:MK表达增高在儿童ALL中可能为预后良好的指标.     

骨髓增生异常综合征患者骨髓CD117的表达及意义

目的 探讨骨髓增生异常综合征(MDS)患者骨髓CD117的表达及意义.方法 采用直接免疫荧光标记法标记细胞表面分化抗原,经流式细胞术测定,对31例MDS患者骨髓的CD117表达及意义进行分析.依据MDS的WHO分型方案、染色体核型以及国际预后积分系统(IPSS)将MDS患者划分为RA/RARS/RCMD组、RAEB Ⅰ/RAEBⅡ组;染色体良好组、染色体不良组;中危Ⅰ组、中危Ⅱ组、高危组.结果 (1)RAEB Ⅰ/RAEB Ⅱ组CD117表达阳性率及表达荧光强度均较RA/RARS/RCMD组高(P<0.01);(2)染色体不良组CD117表达阳性率明显高于染色体良好组(P<0.05);(3)CD117在高危组、中危Ⅱ组的表达阳性率较中危Ⅰ组高(P<0.05),且CD117在高危组、中危Ⅰ组,中危Ⅱ组与中危Ⅰ组间的表达差异亦有显著统计学意义(P均<0.01).结论 CD117检测有助于MDS的分型及预后判断.     

血清降钙素原在老年性肺炎诊断中的应用价值

目的探讨血清降钙素原在老年性肺炎诊断中的应用价值。方法选取2016年5月至2017年4我院266例老年性肺炎患者设为研究组,并根据CURB-65评分将其分为低危组73例、中危组116例及高危组77例,同时选取同期来我院治疗的87例非感染性支气管疾病患者作为对照组,所有患者于治疗前后、治愈或死亡前检测血清降钙素原(PCT)及C反应蛋白(CRP)水平,比较其检测结果。结果治疗前研究组患者的PCT、CRP水平均显著高于对照组,两组比较差异有统计学意义(P<0.05),血清PCT水平随肺炎严重程度的增加而增高,各小组CRP水平比较无显著性差异(P>0.05),治疗后5 d低危组和中危组的PCT水平较治疗前显著下降(P<0.05),高危组治疗前后PCT水平比较无显著性差异(P>0.05),治疗前后各组CRP水平比较无显著性差异(P>0.05),治愈患者PCT≤10 ng/mL的占比显著高于死亡患者,PCT>10 ng/mL的占比显著低于死亡患者,二者比较差异有统计学意义(P<0.05)。结论血清降钙素原可有效的诊断老年性肺炎,并对其严重程度及预后进行判断,其敏感性显著高于C反应蛋白,对老年性肺炎的诊断具有重要的参考价值。     

PHLPP phosphatases as a therapeutic target in insulin resistance-related diseases

Introduction: Pleckstrin homology domain leucine-rich repeat protein phosphatases (PHLPPs), originally identified as Akt kinase hydrophobic motif specific phosphatases, have subsequently been shown to regulate several molecules recurring within the insulin signaling pathway. This observation suggests that PHLPP phosphatases may have a clinically relevant role in the pathogenesis of insulin resistance-related diseases and may thus represent suitable targets for the treatment of these conditions.

Areas covered: The literature pertaining to PHLPPs substrates is reviewed herein, along with information on the molecular players involved in regulating the activity and expression of PHLPP phosphatases. In the present review, knowledge of genetic variants in the genes that encode for PHLPP isozymes and the surrounding regulatory regions is also summarized. In addition, data from the studies addressing the role of PHLPPs in insulin resistance-related disorders and from those investigating the possibility to manipulate these phosphatases for therapeutic purposes are presented.

Expert opinion: A number of issues should be resolved before PHLPPs are pursued as therapeutic targets including: the mechanisms regulating the specificity of PHLPP isozymes; the possibility of differentially regulating PHLPP family members and the possible impact of PHLPPs modulation on the risk of cancer.     

Role of PHLPP1 in inflammation response: Its loss contributes to gliomas development and progression

PH domain leucine-rich repeats protein phosphatase 1(PHLPP1) belongs to a novel family of Ser/Thr protein phosphatases: PHLPP serves as tumor suppressor in several cancers. However, little knowledge about the expression of PHLPP1 in human glioma tumor tissue and its role in inflammation response in glioma cells was known. Glioma samples were obtained from a total of 37 patients including 16 males and 21 females with surgical removal of the brain tumor. PHLPP1 protein and inflammatory cytokines were measured by Western blot analysis and immunohistochemistry while mRNA was determined by RT-PCR. The levels of inflammatory cytokines including TNF-α, IL-17, IL-1β in U251 glioma cells were evaluated by siRNA PHLPP1 and PHLPP1 addition. The loss of PHLPP1 expression occurs at high frequency in human gliomas. The highest mean values of PHLPP1 mRNA and protein were found in non-glioma brain tissues whereas the lowest mean values were found in those in glioblastoma with an increase of TNF-α, IL-17, IL-1β (p < 0.05). PHLPP1 expression in human glioma was associated negatively with the severity of the tumor and inflammatory cytokines. siRNA PHLPP1 could increase the levels of inflammatory cytokines in U251 glioma cells while PHLPP1 addition could inhibit significantly inflammatory cytokines. We concluded that PHLPP1 played a suppression role in inflammatory response of glioma. The present study indicated that PHLPP1 could be used as a predictor for the prediction of the patients or as a therapeutic target for the treatment of human glioma.     

MiR-199a-5p and miR-375 affect colon cancer cell sensitivity to cetuximab by targeting PHLPP1

Objectives: We aimed to analyze the differentially-expressed miRNAs in colon cancer cells in order to identify novel potential biomarkers involved in cancer cell resistance.

Design and methods: We investigated the miRNA expression profile of GEO human colon carcinoma cells, sensitive to the EGFR inhibitor Cetuximab (CTX) and their CTX-resistant counterpart (GEO CR) by using a miRNA chip.

Results: We found 27 upregulated and 10 downregulated miRNAs in GEO CR compared with GEO cells with a fold change ≥ 2. Among the upregulated miRNAs, we focused on miR-199a-5p and miR-375. We report that their enforced expression promotes CTX resistance, whereas their silencing sensitizes to the same drug. The ability of miR-199a-5p and miR-375 to target PHLPP1 (PH domain and leucine-rich repeat protein phosphatase 1), a tumor suppressor that negatively regulates the AKT pathway, accounts, at least in part, for their drug-resistance activity. Indeed, restoration of PHLPP1 increases sensitivity of the GEO cells to CTX and reverts the resistance-promoting effect of miR-199a-5p and miR-375.

Conclusion: This study proposes miR-199a-5p and miR-375 as contributors to CTX resistance in colon cancer and suggests a novel approach based on miRNAs as tools for the therapy of this tumor.     

MicroRNA-302a promotes neointimal formation following carotid artery injury in mice by targeting PHLPP2 thus increasing Akt signaling

The excessive proliferation and migration of smooth muscle cells(SMCs)play an important role in restenosis following percutaneous coronary interventions.MicroRNAs are able to target various genes and involved in the regulation of diverse cellular processes including cell growth and proliferation.In this study we investigated whether and how MicroRNAs regulated vascular SMC proliferation and vascular remodeling following carotid artery injury in mice.We showed that carotid artery injury-induced neointimal formation was remarkably ameliorated in microRNA(miR)-302 heterozygous mice and SMC-specific miR-302 knockout mice.In contrast,delivery of miR-302a adenovirus to the injured carotid artery enhanced neointimal formation.Upregulation of miR-302a enhanced the proliferation and migration of mouse aorta SMC(MASMC)in vitro by promoting cell cycle transition,whereas miR-302a inhibition caused the opposite results.Moreover,miR-302a promoted Akt activation by corporately decreasing Akt expression and increasing Akt phosphorylation in MASMCs.Application of the Akt inhibitor GSK690693(5μmol/L)counteracted the functions of miR-302a in promoting MASMC proliferation and migration.We further revealed that miR-302a directly targeted at the 3′untranslated region of PH domain and leucine rich repeat protein phosphatase 2(PHLPP2)and negatively regulated PHLPP2 expression.Restoration of PHLPP2 abrogated the effects of miR-302a on Akt activation and MASMC motility.Furthermore,knockdown of PHLPP2 largely abolished the inhibition of neointimal formation that was observed in miR-302 heterozygous mice.Our data demonstrate that miR-302a exacerbates SMC proliferation and restenosis through increasing Akt signaling by targeting PHLPP2.     

Degraded and undegraded carrageenans and experimental gastric and duodenal ulceration

The prevention of histamine-induced gastric and duodenal ulceration in the guinea-pig has been examined using a series of undegraded and degraded carrageenans. Undegraded carrageenans were active at lower doses than degraded carrageenans. The high viscosity of the undegraded carrageenans in solution prevented their use in larger doses. Degradation of carrageenan without serious loss of sulphate, gives a product which allows the dose to be increased to an extent that its effect more than offsets the slight loss in activity caused by the degradation. No single feature of carrageenan structure can be related to anti-ulcer activity although degradation, and hence reduction of molecular size, generally reduces activity. Sulphate contents over 30% have little apparent effect on activity; κ-carrageenans were not consistently different in anti-ulcer activity from Λ-carrageenans. This contrasts with the antipeptic activity of carrageenans where κ-carrageenans are less active than their Λ-counter-parts. As with antipeptic activity, the degree of anti-ulcer activity is probably determined by a combination of structural features which includes molecular size and polyanionic properties.