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人附睾蛋白4在早期子宫内膜癌中的诊断价值

人附睾蛋白4在早期子宫内膜癌中的诊断价值杨淑丽;苗劲蔚;何玥;李巍;吴玉梅【摘要】目的探讨人附睾蛋白4(HE4)在子宫内膜腺癌中的诊断价值.方法选取2014年5～12月首都医科大学附属北京妇产医院收治的病理证实的36例Ⅰ期子宫内膜癌患者(子宫内膜癌组)、19例子宫内膜非典型增生患者(子宫内膜非典型增生组)及45例正常子宫内膜患者(正常子宫内膜组)为研究对象,采用酶联免疫吸附试验法分别检测各组血清HE4水平,并对其诊断价值进行分析.结果子宫内膜癌组HE4水平[(65.01 ±40.54) pmol/L]显著高于子宫内膜非典型增生组[(41.01±15.23) pmol/L]及正常子宫内膜组[(40.03±19.03) pmol/L],差异均有高度统计学意义(P＜0.01);正常子宫内膜组与子宫内膜非典型增生组HE4水平比较差异无统计学意义(P＞ 0.05);不同肌层浸润深度、组织分化程度及脉管间隙受浸程度的子宫内膜癌患者血清HE4水平不同(P＜0.05);受试者工作特征曲线下面积AUC 为0.733.根据ROC曲线确定HE4最佳临界值为46.25 pmol/L,该临界值早期诊断子宫内膜癌的敏感性为68.4％,特异性为76.6％.结论早期子宫内膜癌患者血清HE4水平明显升高,可作为子宫内膜癌早期诊断的重要指标,并与肌层浸润深度、组织分化及脉管间隙受浸相关,可用于术前高危因素的预测及评估.%Objective To explore the diagnostic value of serum human epididymis protein 4 (HE4) in early endometrial carcinoma.Methods Thirty-six cases of stage Ⅰ endometrium carcinoma (endometrial carcinoma group),19 cases of en dometrium atypical hyperplasia (endometrium atypical hyperplasia group) and 45 cases of normal endometrium (normal endometrium group) from May to December 2014 in Bejing Obstetrics and GynecologyHospital,Capital Medical University were selected,the levels of serum HE4of patients in each group were detected by enzyme-linked immunosorbent assay (ELISA).The diagnostic value of serum HE4 levels wereanalyzed.Results The level of HE4 in endometrial car cinoma group [(65.01±40.54) pmol/L] was significantly higher than that in endometrium atypical hyperplasia group [(41.01±15.23) pmol/L] and the normal endometrium group [(40.03±19.03) pmol/L],the differences were statistically significant (P ＜ 0.01).There was no statistically significant difference in serum level of HE4 between the endometrium atypical hyperplasia group and endometrium atypical hyperplasia group (P ＞0.05).In endometrial carcinoma group,the levels of serum HE4 were different in patients with different Myometrial infiltration depth,degree of tissue differentiation and vascular clearance.The AUC under the ROC curve was 0.733,and according to the ROC curve the cut-off of HE4 was 46.25 pmol/L,the sensitivity of early diagnosis of endometrial cancer was68.4％,the specificity was 76.6％.Conclusion The level of serum HE4 in Endometial carcinoma patients is significantly increased,it can be used as biological markers of early diagnosis of endometrial carcinoma,the serum level of HE4 may assist to predict and evaluate risk factors before operation.【期刊名称】《中国医药导报》【年(卷),期】2018(015)009【总页数】4页(P70-73)【关键词】子宫内膜肿瘤;人附睾蛋白4;肿瘤标记;诊断【作者】杨淑丽;苗劲蔚;何玥;李巍;吴玉梅【作者单位】首都医科大学附属北京妇产医院妇瘤科,北京 100006;首都医科大学附属北京妇产医院妇瘤科,北京 100006;首都医科大学附属北京妇产医院妇瘤科,北京 100006;首都医科大学附属北京妇产医院妇瘤科,北京 100006;首都医科大学附属北京妇产医院妇瘤科,北京 100006【正文语种】中文【中图分类】R737.3子宫内膜癌是最常见的妇科恶性肿瘤之一，占女性生殖道恶性肿瘤的20%~30%，近年来发病率在世界范围内呈上升趋势[1]。

人附睾蛋白4(HE4)对子宫内膜癌诊断准确性研究

人附睾蛋白4（HE4）对子宫内膜癌诊断准确性研究人附睾蛋白4（HE4）对子宫内膜癌诊断准确性研究引言：子宫内膜癌是女性生殖系统最常见的恶性肿瘤之一，早期诊断对于提高治疗效果和预后至关重要。

人附睾蛋白4（human epididymis protein 4, HE4）作为一种新的肿瘤标志物，被广泛研究其在子宫内膜癌诊断中的准确性。

方法：我们回顾性分析了100例经病理证实的子宫内膜癌患者和100例健康对照者的血清样本。

使用ELISA法检测血清中HE4的表达水平，并将结果与病理诊断结果进行比较。

同时，应用ROC曲线分析计算HE4的敏感性、特异性、阴性预测值（NPV）和阳性预测值（PPV）。

结果：我们发现，子宫内膜癌患者的血清HE4表达显著高于健康对照组（P<0.001）。

根据ROC曲线分析，HE4的诊断准确性良好，敏感性为83%，特异性为79%，NPV为85%，PPV为77%。

此外，将HE4与CA125联合检测在子宫内膜癌的诊断中显示出较好的组合准确性，敏感性为88%，特异性为81%，NPV为89%，PPV为80%。

讨论：凭借其在子宫内膜癌诊断中的较高敏感性和特异性，HE4已成为一个潜在的肿瘤标志物。

与传统的肿瘤标志物CA125相比，HE4具有更好的特异性，这对于筛选高危人群并辅助早期诊断非常重要。

通过组合检测HE4和CA125，可以进一步提高诊断准确性。

然而，尽管HE4在子宫内膜癌诊断中显示出潜力，但其作为独立标志物的应用还需要更多的研究验证。

结论：本研究证实了HE4在子宫内膜癌诊断中的潜力。

HE4可以作为一个辅助标志物，提高子宫内膜癌的早期筛查和诊断准确性。

进一步的研究还需要在更大的病例组中进行，以验证HE4作为独立标志物的可行性，并建立HE4的最佳阈值和诊断模型。

该研究的结果对于提高子宫内膜癌的早期诊断和改善治疗效果具有重要意义，有望为临床实践提供指导，为患者提供更好的预后。

然而，需要更多的临床研究来验证和完善这一领域的相关指南和标准，以使HE4在临床应用中发挥更大的作用综合上述结果，HE4作为一个潜在的肿瘤标志物在子宫内膜癌的诊断中显示出较高的敏感性和特异性。

联合应用人附睾蛋白4与卵巢恶性肿瘤风险算法诊断卵巢癌的研究进展

联合应用人附睾蛋白4与卵巢恶性肿瘤风险算法诊断卵巢癌的研究进展肖林;郭梅;李凤焕;杜丽敏【摘要】Ovarian cancer is a common malignant tumor in women, and its incidence is increasing year by year. Because there is no specific clinical manifestation in the early stage of the disease, the early diagnosis of ovarian cancer needs the help of imaging and serological indicators. Human epididymis protein 4 (HE4) is a new type of serum tumor markersin recent years, compared with the traditional ovarian cancer tumor markers CA125, its sensitivity and specificity have a greater degree of improvement. However, due to the interference of many factors on theHE4 level, the concept of risk of ovarian malignancy algorithm (ROMA) was proposed through the inclusion of CA125, HE4 and other levels of the comprehensive assessment of early ovarian cancer. In this paper, the research progress of HE4 and ROMA in the diagnosis of ovarian cancer in recent years is reviewed.%卵巢癌是女性较为常见的一种恶性肿瘤，其发病率正呈现逐年升高的趋势。

人附睾蛋白4在子宫内膜癌诊断中的临床价值

关，血清ＨＥ４的检测有助于对子宫内膜癌症的辅助诊断。
关键词：人附睾蛋白４；子宫内膜癌；子宫内膜增生
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子宫内膜癌患者术前和术后的血清ＨＥ４浓度，探讨ＨＥ４对子
宫内膜癌的辅助诊断价值。
ｌ资料与方法
后比较； △：Ｐ＜Ｏ．０５，与子宫内膜增生组比较。
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年龄比较，差异无统计学意义（Ｐ＞Ｏ．０５）。１．２方法清晨用普通真空试管采集所有研究对象空腹静脉血３ｍｌ，室温静置１ｈ左右，以３０００ｒ／ｍｉｎ离心１５ｍｉｎ，分离
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人附睾蛋白４在子宫内膜癌诊断中的临床价值

人附睾分泌蛋白4在子宫内膜癌组织中的表达及临床意义

人附睾分泌蛋白4在子宫内膜癌组织中的表达及临床意义目的：通过检测人附睾分泌蛋白4（HE4）在子宫内膜癌组织中的表达水平，观察HE4与不同临床及病理特征的相关性，探讨其在子宫内膜癌诊断、治疗及预后中的临床意义。

方法：应用即用型免疫组织化学染色两步法对32例经病理学证实为子宫内膜癌的石蜡标本进行HE4检测。

分析HE4表达与子宫内膜癌FIGO分期、病理类型、组织学分级、肌层侵犯深度、肿瘤的大小和年龄的关系等各组间的差异，分析其与子宫内膜癌临床病理特征的关系。

结果：（1）HE4阳性颗粒主要定位在细胞膜和细胞质上，在32例子宫内膜癌组织中的HE4的阳性表达率较高，为87.5%。

（2）HE4在组织学分级为高分化组的子宫内膜癌组织中的阳性表达率及强度均高于中低分化组（P＜0.05）。

（3）HE4蛋白在晚期子宫内膜癌组织中阳性表达率及表达强度均高于早期组（P＜0.05）。

（4）HE4蛋白在子宫内膜癌的阳性表达与年龄、病理分型、肌层侵犯深度及肿瘤大小均无关（P>0.05）；但在HE4强阳性表达程度方面，深肌层浸润者强阳性表达（61.5%）多于无深肌层浸润者（36.8%），其直径≥2 cm的肿瘤强阳性表达（61.1%）高于直径4分为阳性，>6分为强阳性。

病理切片分别由3位高年资病理医师轮流读片评定。

1.4 统计学处理运用SPSS 18.0统计学软件对HE4在不同子宫内膜癌组织中的表达差异进行相关性分析，计量资料用（x±s）表示，比较采用t检验；计数资料以率（%）表示，比较采用字2检验，P＜0.05为差异有统计学意义。

2 结果2.1 HE4在子宫内膜癌组织中的表达HE4阳性颗粒主要定位在细胞膜及细胞浆上；着色呈棕黄色或棕褐色；视野内阳性细胞呈点、片状分布，见图1～5。

在32例癌组织中的HE4阳性13例（阳性表达率为40.6%），强阳性15例（阳性表达率为46.9%），总阳性表达率为87.5%。

2.2 HE4在不同临床病理特征的子宫内膜癌组织中的表达2.2.1 子宫内膜癌组织中HE4表达与组织学分级的关系HE4在组织学分级分别在高、中-低分化组的子宫内膜癌组织中的阳性表达率为62.5%、95.8%，两者比较差异有统计学意义（字2=8.253，P=0.004）。

人附睾蛋白4在子宫内膜癌诊断与治疗中的研究进展

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带有硬物或尖物，在治疗摆位时直接刺破真空袋而导致漏气。在真空袋使用过程中除了质量把关外，操作人员应尽量
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气原因： ①不同厂家真空袋质量差距较大，对真空袋漏气现象有着直接的影响。② 真空袋的真空阀门密封不严或治疗过程中阀门松动，是真空袋漏气的最主要原因。③ 患者身上

A Competitive Infection Model of Hematogenously Disseminated

A Competitive Infection Model of Hematogenously Disseminated Candidiasis in Mice Redeﬁnes the Role of Candida albicans IRS4in PathogenesisSuresh B.Raman,a M.Hong Nguyen,a Shaoji Cheng,a Hassan Badrane,a Kenneth A.Iczkowski,c Marilyn Wegener,b Sarah L.Gaffen,a Aaron P.Mitchell,d Cornelius J.Clancy a,bDepartment of Medicine,University of Pittsburgh,Pittsburgh,Pennsylvania,USA a;VA Pittsburgh Healthcare System,Pittsburgh,Pennsylvania,USA b;Department of Pathology,University of Colorado,Denver,Colorado,USA c;Department of Biological Sciences,Carnegie Mellon University,Pittsburgh,Pennsylvania,USA dCandida albicans IRS4encodes a protein that regulates phosphatidylinositol-(4,5)-bisphosphate,which was shown to contrib-ute to hematogenously disseminated candidiasis(DC)after several days in the standard mouse model.Our objective was to more accurately deﬁne the temporal contributions of IRS4to pathogenesis.During competition assays in vitro,an irs4-null(⌬irs4) mutant exhibited wild-typeﬁtness.In DC experiments,mice were infected intravenously with the⌬irs4mutant,strain CAI-12 (1؋105CFU),or a mixture of the strains(0.5؋105CFU each).In single-strain infections,quantitative PCR revealed reduced ⌬irs4mutant burdens within kidneys at days1,4,and7but not6h.In competitive infections,the⌬irs4mutant was outcom-peted by CAI-12in each mouse at>6h(competitive indices,P<0.0001).At4and7days,the⌬irs4mutant burdens during com-petitive infections were signiﬁcantly lower than those during single-strain infections(P؍0.01and P<0.001,respectively),sug-gesting increased susceptibility to inﬂammatory responses.Phagocytic inﬁltration of kidneys in response to CAI-12or competitive infections was signiﬁcantly greater than that in response to⌬irs4mutant infection at days1and4(P<0.001),and the⌬irs4mutant was more susceptible to phagocytosis and killing by human polymorphonuclear cells(P؍0.01and P؍0.006, respectively)and mouse macrophages in vitro(P؍0.04and P؍0.01,respectively).Therefore,IRS4contributes to tissue inva-sion at early stages of DC and mediates resistance to phagocytosis as DC progresses.Microarray analysis revealed remarkably similar gene expression by the⌬irs4mutant and reference strain CAI-12within blood,suggesting that IRS4is not signiﬁcantly involved in the hematogenous stage of disease.A competitive DC model detects attenuated virulence that is not evident with the standard model.C andida albicans is the major fungal pathogen among hospital-ized patients in the developed world,causing a wide range of superﬁcial,mucosal,and invasive infections(1).Candidemia is the fourth most common bloodstream infection in the United States and is associated with mortality rates approaching or ex-ceeding40%,despite antifungal therapy(2,3).The development of new treatment,prevention,and diagnostic strategies against candidemia and disseminated candidiasis depends upon better understanding of the pathogenesis of C.albicans infections.In previous studies,we demonstrated that C.albicans IRS4was essential for full virulence during hematogenously disseminated candidiasis in mice.IRS4encodes an Eps15homology(EH)do-main protein that physically interacts with the5=-phosphatase Inp51to regulate the levels and plasma membrane distribution of phosphatidylinositol-(4,5)-bisphosphate[PI(4,5)P2](4–6).Dis-ruption of IRS4resulted in cell wall derangements and impaired hyphal formation within solid agar and mouse kidneys.The tissue burdens of the irs4-null(⌬irs4)mutant within mouse kidneys were similar to those of wild-type C.albicans after20h.However, the⌬irs4mutant was unable to form dense mats of hyphae,and tissue burdens at4days were signiﬁcantly attenuated(4,5).The data suggested that PI(4,5)P2regulation is required for the pro-gression but not initiation of kidney infection.We hypothesized that the attenuated virulence of the⌬irs4mutant results from a failure to maintain cell wall integrity in the face of ongoing cell wall stress during tissue invasion.At the same time,we recognized that IRS4may make contributions to pathogenesis within the kid-neys that were not apparent in the mouse model.Indeed,mutants were signiﬁcantly impaired in adherence to various epithelial cell lines in vitro,suggesting that IRS4is relevant at the early stages of tissue invasion.Furthermore,if the impaired cell wall integrity of the⌬irs4mutant was contributing to attenuated virulence at later time points,it was not clear why such defects would not also be important at early time points.In assessing the contribution of IRS4to virulence,we used the standard mouse model of hematogenously disseminated candidi-asis(7).In this model,C.albicans strains are inoculated via the lateral tail vein,and the kidneys are the primary target organ. Groups of mice are typically infected with a wild-type,isogenic gene disruption,or gene reinsertion strain,and endpoints like mortality and tissue burdens are compared between groups at serial time points(7).The model has advantages of simplicity and reproducibility,but it is limited by relatively insensitive end-points.Moreover,large numbers of mice are typically required toReceived26July2012Returned for modiﬁcation13August2012Accepted12February2013Published ahead of print19February2013Editor:G.S.Deepe,Jr.Address correspondence to Cornelius J.Clancy,cjc76@.Supplemental material for this article may be found at /10.1128/IAI.00743-12.Copyright©2013,American Society for Microbiology.All Rights Reserved.doi:10.1128/IAI.00743-12 Infection and Immunity p.1430–1438May2013Volume81Number5 on January 18, 2015 by guest / Downloaded fromdemonstrate that virulence is attenuated in mutant strains,al-though expenditure of lives may be reduced by the technical skill of investigators and in cases of large differences between strains. Competitive infection models,in which individual hosts are in-fected with a mixture of microbial strains,are powerful tools for identifying small differences inﬁtness(i.e.,relative virulence)be-tween strains(8–16).By eliminating interhost variability,they achieve high levels of sensitivity and spare animals.In recent years, investigators have begun to use competitive models of oral,gas-trointestinal,and disseminated candidiasis in lieu of conventional mouse models to compare the virulence of wild-type and mutant C.albicans strains(8,10,11,13,15,16).These studies have con-clusively identiﬁed C.albicans genes that encode virulence deter-minants,but they have not tapped the full potential of competitive models to characterize the pathogen-host interaction.In particu-lar,the studies have not compared C.albicans strain behavior and host responses during both competitive and single-strain infec-tions,which is a particularly useful strategy for deﬁning the con-tribution of a speciﬁc gene to the pathogenic process over the time course of disease.The primary objective of the present study was to deﬁne more accurately the temporal contribution of C.albicans IRS4to the pathogenesis of disseminated candidiasis.Toward this end,we developed a competitive infection model of hematogenously dis-seminated candidiasis in mice and characterized gene expression by the⌬irs4mutant within blood.We hypothesized that these methods would demonstrate that the virulence of the⌬irs4mu-tant is attenuated at the early stages of kidney invasion,rather than after several days,as suggested in our prior studies. MATERIALS AND METHODSC.albicans strains and growth conditions.C.albicans⌬irs4and an IRS4 reinsertion strain were created and characterized as described in our pre-vious publications(4,5)(Table1).C.albicans CAI-12was the IRS4-intact, isogenic strain used for comparisons throughout the study.Growth rates in vitro were determined in yeast peptone dextrose(YPD)and Sabouraud dextrose(SD)media at30and37°C in microtiter plates,as described previously(19).Extended growth curves in vitro were assessed through four dilution and regrowth cycles of24h each.To induce hyphal forma-tion in liquid media,C.albicans strains grown overnight on YPD agar were subcultured into liquid YPD supplemented with5%fetal calf serum (FCS)and liquid RPMI1640at37°C(4,5).In preparation for intravenous challenge of mice,all strains were grown to stationary phase overnight in YPD medium at35°C.C.albicans cells were washed in sterile saline,and inocula were prepared at the desired concentration in sterile saline.Quantitative real-time PCR.For the standard curves relating changes in threshold cycle(⌬C T)values to C.albicans cell numbers,DNA samples were prepared from sterile saline or naive kidneys spiked with cells from a mid-logarithmic-phase culture of C.albicans CAI-12grown at30°C in YPD medium.The cell density of the undiluted culture was determined by direct counting with a hemocytometer and veriﬁed by plating.The culture was serially diluted in YPD medium.In both spiking and disseminated candidiasis experiments,kidneys were homogenized and subjected to lyti-case treatment(30min at37°C).Genomic DNA was isolated using aDNeasy blood and tissue kit(Qiagen).Our analyses included at least threebiological replicates for all samples,each from independent collections.The quantitative real-time PCR was performed using PerfeCTa SYBRgreen Fast Mix(Quanta Biosciences Inc.)with an ABI7500sequencedetection system(Applied Biosystems).For each20-␮l PCR mixture,2␮lgenomic DNA was used with6␮l nuclease-free water,10␮l PerfeCTaSYBR green Fast Mix,and1␮l of each primer.Gene-speciﬁc primers forIRS4detection in CAI-12(primers IRS-qPCR-For[5=-ACC AGC AATCTT CCA CTG AGA TCA ACA-3=]and IRS-qPCR-Rev[5=-CTT CCATGG CTT CAA CTC ATT AAA CCT TGA-3=],yielding a138-bp product)and primers speciﬁc to the⌬irs4mutant(primers⌬irs-qPCR For[5=-CTG AGG TGG AAG TTG GAG AAA CAA CC-3=]and⌬irs-qPCR-Rev[5=-CCC TGA TTG ACT GGA ACA GGA TCC TC-3=],yielding a122-bpproduct)were designed.The thermocycler program was95°C for10minand40cycles of95°C for30s and58°C for45s.Quantitative PCR(qPCR)assays were run according to the manufacturer’s directions,and resultswere analyzed with sequence detection system software(v2.0.1;AppliedBiosystems).Each sample was assigned a C T value,which identiﬁes thecycle number during PCR whenﬂuorescence exceeds a threshold valuedetermined by the software.Differences in DNA recovery between sam-ples were normalized by determining the total DNA concentration of eachinfected kidney sample([DNA]sample),comparing that concentration tothe DNA concentration values from uninfected kidney([DNA]uninfected),and adjusting each sample C T value according to the formula C T ad-justedϭC Tϩx,where x is log2([DNA]sample/[DNA]uninfected)(20).⌬C T values were used to calculate genomic equivalents(GE)from a standardcurve generated from DNA samples prepared from uninfected kidneysspiked individually with CAI-12,the⌬irs4mutant,and a reinsertionstrain yeast.All qPCR results for samples from infected tissues are ex-pressed as GE per gram equivalent of tissue.Since a full copy of IRS4wasreintroduced into a disrupted native locus in the reinsertion strain,bothwild-type(IRS4)-speciﬁc and⌬irs4mutant-speciﬁc primers ampliﬁedsignals.Therefore,in the mixed infection(infection with CAI-12plus thereinsertion strain),the IRS4primer set ampliﬁed both wild-type allelesfrom CAI-12and one wild-type allele from the reinsertion strain(wild-type total GE),and the⌬irs4mutant primer set ampliﬁed one allele fromthe reinsertion strain(reinsertion strain GE).The actual number of wild-type GE in the mixed infection was calculated using the formula wild-type total GEϪreinsertion strain GE.No signal was detected when thefollowing templates were tested as negative controls:genomic DNA pre-pared from kidneys of uninfected mice,mutant primers for kidneysspiked or infected with CAI-12,and CAI-12primers for kidneys spiked orinfected with the⌬irs4mutant.Competitive infection model.Mouse experiments were approved bythe Institutional Animal Care and Use Committee(IACUC)at the Uni-versity of Pittsburgh.Groups of7-week-old male ICR mice(8to10miceper group;Harlan-Sprague)were individually inoculated by single intra-venous injections of the lateral tail vein with200␮l of sterile saline con-taining(i)1ϫ105CFU of CAI-12,the⌬irs4mutant,or the IRS4reinser-tion strain;(ii)0.5ϫ105CFU each of CAI12and the⌬irs4mutant,or(iii)0.5ϫ105CFU each of CAI-12and the reinsertion strain.Mice wererandomly selected to be included in a given group.Strain concentrationswere conﬁrmed by serial dilution of inocula and enumeration of CFU.Mice in the CAI-12,the⌬irs4mutant,and the CAI-12–⌬irs4mutantgroups were sacriﬁced at6h,1day,4days,and7days after intravenousinoculation,and their kidneys were aseptically removed.Reinsertionstrain-infected groups were sacriﬁced on days1and4after challenge.C.albicans CAI-12does not kill mice at this inoculum over7days,whichensured that interpretation of tissue burdens would not be obscured bydeath.Kidneys were sectioned such that one-half of the left kidney andone-half of the right kidney were combined and used for CFU determi-nation,while the other halves were combined and used for qPCR analysis.The organs were weighed and homogenized in2ml sterile phosphate-TABLE1C.albicans strainsStrain Description ReferenceSC5314Clinical isolate17CAI-12a⌬ura3::imm434/URA318⌬irs4mutant a⌬irs::hisG/⌬irs::hisG⌬ura3::imm434/URA34IRS4reinsertion a⌬irs::hisG/IRS::URA3-hisG⌬ura3::imm434/⌬ura3::imm4344a Strains did not differ in activity of orotidine5=-monophosphate decarboxylase.Competitive C.albicans Infections in Mice1431 on January 18, 2015 by guest / Downloaded frombuffered saline(PBS).For CFU enumeration,serial dilutions were plated onto SD plates containing piperacillin(60mg mlϪ1)and amikacin(60mg mlϪ1).The plates were incubated at30°C for48h.Values were expressed as the log number of CFU per gram kidney.Differences in tissue burdens (numbers of GE or CFU)between strains were compared using analysis of variance(ANOVA).Pairwise comparisons were calculated using a Bon-ferroni adjustment.Within individual mice during competitive infec-tions,results were expressed as the competitive index(CI).CI was deﬁned as[(log10GE/g kidney for mutant strain/log10GE/g kidney for CAI-12)/ (log10GE/ml inoculum for mutant strain/log10GE/ml inoculum for CAI-12)](9).Results were compared using Student’s t test.Histopathology.Histopathology was performed by a pathologist blinded to the experimental design.Kidneys wereﬁxed with formalin and embedded in parafﬁn,after which thin sections were prepared and stained with hematoxylin-eosin(H&E)or periodic acid-Schiff(PAS)(21).For each strain,kidneys from three mice were chosen for image analysis.TIFF images of all the tissue on each of the slides were captured.The images were analyzed on a Windows XP personal computer using the public domain National Institutes of Health(NIH)image program ImageJ(http: ///nih-image/).For each image,outline splines were traced around the total area(s)of tissues,and another series of outline splines was traced around the area(s)involved in acute inﬂammation(5). At least20images for each kidney were analyzed.The percentage of the total area with inﬂammation was calculated and expressed as the meanϮstandard deviation.Phagocytosis and killing assays.Phagocytosis and killing assays were performed as previously described(19,22),with slight modiﬁcations. Polymorphonuclear cells(PMNs)were isolated from heparinized blood by dextran sedimentation,followed by centrifugation through Ficoll-Hypaque.After removing contaminating erythrocytes by hypotonic lysis, the PMNs were resuspended in RPMI1640.Prior to phagocytosis and killing assays,C.albicans strains were opsonized with50%normal human serum at30°C for30min.For phagocytosis assay,0.5ml of opsonized C. albicans cells was incubated with0.5ml of PMNs at a PMN/C.albicans ratio of1:1in1ml RPMI1640at37°C for15min on a shaker.Three drops of the sample were then cytospun and Gram stained.Percent phagocytosis was calculated as the proportion of PMNs containing one or more yeast cells after counting of100PMNs.For the killing assay,opsonized C.albi-cans strains were incubated with106PMNs at a PMN/C.albicans ratio of 50:1in1ml of RPMI1640containing5%human serum at37°C for2h with gentle shaking.After complete lysis of PMNs with sterile water,serial 10-fold dilutions were made and colony counts were enumerated.For mouse macrophage(J774A.1)studies,cells procured from ATCC were generated in a96-well plate(5ϫ105cells/well)by incubation for30min at37°C in5%CO2.Prior to phagocytosis and killing assays,C.albicans strains suspended in modiﬁed Eagle’s medium(MEM)were opsonized with5%normal human serum at room temperature for30min.For the phagocytosis assay,200␮l opsonized C.albicans cells was added to wells containing the monolayer(effector/target cell ratio,ϳ10:1)and the cells were incubated for15min at37°C in5%CO2.After incubation,the supernatant was aspirated and washed with prewarmed MEM.Serial10-fold dilutions were made,and colony counts were enumerated.For the killing assay,200␮l MEM-Sabouraud broth(1:1)was added to wells containing monolayers with phagocytosed C.albicans cells,and the plates were incubated for3h at37°C in 5%CO2.The monolayers were scraped,and after complete lysis of macro-phages with sterile water,serial10-fold dilutions were made and colony counts were enumerated.The phagocytosis and killing experiments were per-formed in triplicate and repeated at least twice.The percentage of phagocy-tized C.albicans cells was deﬁned as[1Ϫ(number of uningested CFU/num-ber of CFU at the start of incubation)]ϫ100.The percentage of C.albicans cells killed by the phagocytes was deﬁned as[1Ϫ(number of CFU after incubation in the presence of phagocytes/number of CFU phagocytosed)]ϫ100.Thefungicidalactivitywascalculatedasthepercentsurvivalof C.albicans after2h of incubation with PMNs.Assessment of gene expression in whole blood using whole-genome microarrays.C.albicans Genome Oligo Set v1.1(Qiagen,Valencia,CA)was resuspended at30␮M in ArrayIt Micro Spotting Solution Plus(Telechem,San Jose,CA)and printed on UltraGAPS slides(Corning,Corning,NY)using a Genemachine Omnigrid arrayer(Genomic Instru-mentation Services,San Carlos,CA).Each slide had two replicated arrayswith6,936features,including70-mer oligonucleotides representing5,948predicted open reading frames(ORFs),318cloned genes,and random70-mer controls.Transcriptional proﬁling experiments in human bloodwere performed as previously described(23).In brief,three biologicalreplicates of C.albicans CAI-12and the⌬irs4mutant were grown tomid-exponential phase in YPD medium at30°C,washed once in0.1MPBS,pH7.0,and resuspended in this buffer at a density of37.5ϫ108cells/ml.After incubation for30min at37°C,100␮l of the suspension wasinoculated into7.5ml of fresh whole human blood from one of the inves-tigators(C.J.C.),which was collected in a heparinized tube.After incuba-tion of the tube with gentle shaking for2h at37°C,cells were harvestedand mixed with1g of425-to600-␮m-diameter glass beads(SigmaChemical Co.,St.Louis,MO).Total RNA was extracted using an RNeasyminikit(Qiagen)and a Mini-BeadBeater(Biospec Products,Bartlesville,OK),and on-column DNase digestion with an RNase-free DNase set(Qiagen)was systematically performed.Twenty to25␮g of total RNA wasused to generate labeled cDNA with a SuperScript indirect cDNA labelingsystem(Invitrogen,Carlsbad,CA).Two cDNAs from different sampleslabeled with different dyes(Cy3and Cy5)were mixed,denatured in3.4ϫSSC(1ϫSSC is0.15M NaCl plus0.015M sodium citrate),0.3%SDS,0.5␮g/␮l salmon sperm DNA,and hybridized overnight(14to16h)to mi-croarray slides.A dye-ﬂip strategy was employed.After washing and dry-ing,the slides were scanned in a GenePix4000B scanner(Axon Instru-ments,Union City,CA).GenePix Pro v3.0software(Axon)was used toextract the spot intensities for bothﬂuorescent dyes,and data were ar-chived in Microsoft Access software.For individual slides,the thresholdintensity for eachﬂuorescent channel was established as the average in-tensity of spots corresponding to negative-control70-mers plus1.5timesthe standard deviation.Only intensities higher than the threshold wereconsidered.Replicated spots in each slide had to have a coefﬁcient ofvariation in intensities ofϽ65%.The intensities of the replicated spots ineach slide that passed the criterion were averaged and used for normaliza-tion.We performed a global normalization among all slides so that thetotal slide intensity for each channel was identical among slides and chan-nels.The ratio of the intensity of the⌬irs4mutant to that of strain CAI-12was used for further analysis.We applied signiﬁcance analysis of microarrays(SAM)to detect dif-ferentially expressed ORFs(24).SAM assigns a score to each gene on thebasis of the change in gene expression relative to the standard deviation ofrepeated measurements.For genes with scores greater than an adjustablethreshold,SAM uses permutations of the repeated measurements to esti-mate the percentage of genes identiﬁed by chance,the false discovery rate(FDR).SAM has been shown to be superior to conventional microarrayanalysis based on pairwise fold change methods.Cluster software andTreeview software were used for cluster analysis and display of genome-wide expression data(25).Differentially expressed ORFs were assignedgene names,descriptions,and pathways on the basis of assignments in theCandida Genome Database(/).Gene expression by the strains was conﬁrmed for eight genes by re-verse transcription-PTR(GCA2,ERG16,RNR22,CWH8,SKN1,CHT2,RHO1,and ERK1),using primers designed and synthesized by Invitrogen(see Table S1in the supplemental material).cDNA was synthesized using1␮g total RNA,2.5␮M reverse primers,500␮M deoxynucleosidetriphosphate(dNTP)mix,5mM dithiothreitol,40U RNaseOUT,1ϫﬁrst-strand buffer,and100U SuperScript III reverse transcriptase(Invit-rogen)at46°C for2h.Ampliﬁcation was performed for30cycles,whichgave band intensities linearly proportional to the amount of cDNA.Thereaction mixtures included1␮l a1/10dilution of cDNA in0.2mM dNTPmix,1.5mM MgCl2,0.2␮M each primer,1ϫPCR buffer,and1.5UPlatinum Taq DNA polymerase(Invitrogen).Ten microliters of each PCRRaman et al. Infection and Immunity on January 18, 2015 by guest / Downloaded fromproduct was run in a1.5%agarose gel,and the gel was stained with ethidium bromide.Images were digitalized using a transilluminator con-nected to a camera and Quantity One software(Bio-Rad).The software esti-mated the band intensity for each PCR product,which was used to calculate the ratio of⌬irs4mutant expression/CAI-12expression for each ORF.Statistics.For comparisons across groups(see Fig.1,4,and5), ANOVA was used.Pairwise comparisons were then made using a Bonfer-roni adjustment.Otherwise,comparisons were made using t test.All anal-yses were performed using STATA v11software(College Station,TX).P values ofϽ0.05were considered signiﬁcant.Microarray data accession numbers.Microarray data were deposited in the NCBI GEO database under accession number GSM967204(https: ///geo/query/acc.cgi?accϭGSM967204). RESULTSFitness of⌬irs4mutant in vitro.To distinguish between C.albi-cans strains in competition assays,we designed qPCR primers that targeted speciﬁc segments of IRS4and the hisG cassette used for gene disruption.In titration experiments,serial dilutions of yeasts of strain CAI-12,the⌬irs4mutant,or the IRS4reinsertion strain were suspended in sterile saline and spiked into explanted,unin-fected mouse kidneys.Genomic DNA extracted from suspensions and spiked kidneys was used as a template for qPCR assays that established standard curves(see Fig.S1in the supplemental ma-terial).The signals from these reactions were linear over at least7 orders of magnitude for each strain(all R2values wereՆ0.96). The limits of detection in suspension and spiked kidneys were at or near1and10yeast cells,respectively.As we showed previously,growth rates of CAI-12and the⌬irs4 mutant were indistinguishable after the strains were indepen-dently inoculated into YPD or SD medium at30°C(see Fig.S2in the supplemental material).The growth curves for a mixed culture of CAI-12and the⌬irs4mutant were similar to those of the indi-vidual strains.At all time points,qPCR conﬁrmed that the con-centrations of the strains were the same whether they were grown alone or in mixed cultures.Similar results were obtained in SD medium at30°C and at37°C(not shown).In addition,there were no signiﬁcant differences in the growth curves of the strains through four dilution and regrowth cycles of24h each.The for-mation of hyphae by the strains in liquid medium was compara-ble,consistent with our previous reports(10,11).Relative virulence of the⌬irs4mutant during hematog-enously disseminated candidiasis.To establish our competitive infection model,we infected groups of mice intravenously with CAI-12alone(1ϫ105CFU/mouse),the⌬irs4mutant alone(1ϫ105CFU/mouse),or a1:1mixture of CAI-12and the⌬irs4mutant (0.5ϫ105CFU/strain/mouse).Using conventional methods for enumerating the CFU within the kidneys,we corroborated our earlierﬁndings for single-strain infections:tissue burdens of the the⌬irs4mutant were diminished compared to those of CAI-12at 4and7days but comparable to those of CAI-12at6h or1day(Fig. 1A).The qPCR method of quantitating genome equivalents(GE) detected absolute candidal burdens during single-strain infections that were approximately1log unit higher than those suggested by the numbers of CFU(Fig.1B).By qPCR,the⌬irs4mutant inoc-ulated alone caused signiﬁcantly lower tissue burdens than CAI-12at1day,as well as4and7days.At6h,there were still no differences in tissue burdens between mice infected with the⌬irs4 mutant and mice infected with CAI-12.In the competitive model,the tissue burdens of the⌬irs4mu-tant and strain CAI-12at6h and1day,as determined by qPCR,were identical to those observed when the strains were given alone (Fig.1B).At days4and7,however,the tissue burdens of the⌬irs4 mutant in the competitive model were signiﬁcantly attenuated compared to those from the⌬irs4mutant single-strain infection. The burdens of CAI-12at each time point were identical whether the strain was given alone or with the mutant.Of note,the tissue burdens of an IRS4reinsertion strain were identical to those of CAI-12in both single-strain and competitive infections,indicat-ing that the attenuated virulence of the⌬irs4mutant was a result of gene disruption(see Fig.S3in the supplemental material).Of course,the real power of competitive infection models comes from comparing strains within individual animals,rather than across groups.In these intrahost analyses,results are ex-pressed as the competitive index,which is the ratio of the mutant to a reference ing the competitive index,the⌬irs4mu-tant was signiﬁcantly attenuated at each time point,including6h, FIG1Recovery of CAI-12and the⌬irs4mutant from mouse kidneys.(A) Tissue burdens for groups of mice were determined by enumerating the CFU.(B)Relative tissue burdens were determined by qPCR measurement of ge-nome equivalents per gram of tissue.In a three-way comparison of the number of CFU of tissue burdens(A)at6h,1day,4days,and7days,P values were0.39, 0.03,0.0001,and0.007,respectively(ANOVA).In a four-way comparison of the number of GE relative to tissue burdens(B)at6h,1day,4days,and7days, P values were0.17,Ͻ0.0001,Ͻ0.0001,andϽ0.0001,respectively(ANOVA). *,PϽ0.001for CAI-12versus the⌬irs4mutant at day4;**,Pϭ0.03for CAI-12versus the⌬irs4mutant at day7;***,PՅ0.001for CAI-12versus the ⌬irs4mutant,in single-strain and mixed infections;****,Pϭ0.01for the⌬irs4 mutant in single-strain infection versus the⌬irs4mutant in mixed infections at day4;*****,PϽ0.001for the⌬irs4mutant in single-strain infection versus the⌬irs4mutant in mixed infections at day7.Mix,tissue burdens(number of CFU)during mixed infections,not distinguishing between strains(A);Mix–CAI-12,relative tissue burdens(GE)of CAI-12during mixed infections;Mix-⌬irs4,relative tissue burdens(GE)of the⌬irs4mutant during mixed infec-tions.Data are means of results for8to10mice per group per time point.Error bars represent standard deviations.Competitive C.albicans Infections in Mice1433 on January 18, 2015 by guest / Downloaded from。

人附睾蛋白4在子宫内膜癌中的研究进展

人附睾蛋白4在子宫内膜癌中的研究进展人附睾蛋白4（HE4）是近年来发现的新型的血清学标志物，目前在卵巢上皮性癌已经得到广泛应用。

近来研究发现，HE4在子宫内膜癌组织中高表达，国内外学者及机构已经就其高敏感性和特异性作为子宫内膜癌的肿瘤标志物开展研究。

现就HE4在子宫内膜癌中的研究进展、临床意义等做一综述。

标签：人附睾分泌蛋白4；肿瘤标记；子宫内膜癌；诊断；预后子宫内膜癌是发生于子宫内膜的一种上皮性恶性肿瘤，是女性生殖道常见的三大恶性肿瘤之一。

随着人类生活水平的提高、寿命的延长及内分泌代谢疾病的增加，近年来有上升且年轻化趋势。

临床上选取以手术为主的综合治疗方案，而综合治疗方案在一定程度上依赖于准确的FIGO分期。

大多数子宫内膜癌可通过疾病发生发展过程的一些症状如绝经后阴道不规则流血及辅助检测B超检查、诊断性刮宫、宫腔镜甚至核磁共振及肿瘤标志物糖类抗原125等可得到早期诊治，总体治愈率高。

但由于一些检查手段的局限性致误诊或漏诊；术前评估不足致治疗方式不规范，如手术范围不足或过度治疗；术后随访不足致术后复发等原因，患者术后存活率低。

对于子宫内膜癌症的早期诊断、检测复发和监测对治疗的反应，临床上需要更敏感无创的筛查方法或肿瘤学标志物。

目前国内外学者对人附睾分泌蛋白（HE4）进行了很多研究，肯定了它在子宫内膜癌中的诊断价值，尤其是在早期子宫内膜癌的诊断以及预测疾病分期方面。

目前认为在所有的被测肿瘤标志物中HE4与子宫内膜癌的相关性最强[1]。

1 HE4的来源与分子基础HE4最早在1991年由Kirchhoff等[2]发现于附睾上皮组织，为酸性小分子分泌型糖蛋白，是乳清酸蛋白（whey acidic protein，WAP）结构域家族蛋白中的一员。

HE4基因又名WFDC2（WAP four-disulfide core domains 2），位于20q12-q1312，全长11 178 kb，编码124个氨基酸的HE4蛋白前体。

IDEXX 4Dx

Screening for vector-borne disease IDEXX 4Dx® Plus Test clinical reference guideWith the IDEXX 4Dx ® Plus T est, a positive result can also be an indication of ticks and and the pathogens they carry. Know more with every resultdetect antibodies to these pathogens When you use the IDEXX 4Dx Plus T est as a screening tool, you maycarried by these ticks Anaplasmaphagocytophilum Borrelia burgdorferi (Lyme disease)Ehrlichia ewingiiEhrlichia canis Anaplasma platysBabesia spp.Rocky Mountain spotted feverEhrlichia chaffeensis TularemiaRocky Mountain spotted fever STARIBartonella spp.Babesia spp.Ehrlichia canis Brown dog tickRhipicephalus sanguineusAmerican dog tickDermacentor variabilisBlack-legged tick (deer tick)Ixodes scapularis Ixodes pacificusLone star tickAmblyomma americanumRocky Mountain spotted fever TularemiaGeographic tickdistribution as of 20213that may also transmit other pathogens and infections to dogs and peopleLyme diseasebacterium Borrelia burgdorferi cases that have mild to severe disease.* S erology is typically used to diagnose Lyme disease. B. burgdorferi Did you know?•D ogs testing positive for antibodies to the C 6 peptide had 43% increased risk of having chronic kidney disease (CKD) compared to seronegative dogs.4• T he C 6 peptide used in the IDEXX 4Dx ® Plus Test and Lyme Quant C 6® Antibody Test does not cross-react with the antibody response to commercially available Lyme vaccines.5 • D ogs with seroreactivity to both B. burgdorferi and Anaplasma phagocytophilum may have two times the risk of developing clinical illness than singularly infected dogs.2Borrelia burgdorferiPrimary vectorsIxodes scapularis or Ixodes pacificus Black-legged tick (deer tick)Pathology• Localizes in tissues of infected dogs • Synovitis (may be subclinical) • Lyme nephritisClinical presentationChronic infection with clinical signs that may present acutely:• Fever, anorexia• Polyarthritis, lameness• Rapidly progressive renal failure • Neurologic syndromesLaboratory abnormalities• Elevated C 6 antibody level ≥ 30 U/mL • May have proteinuria• M ay have IDEXX SDMA ® T est result > 14 µg/dLCKD monitoring• Chemistry panel with SDMA – R ecommended to evaluate forthe development of protein-losing kidney disease• Urinalysis with Reflex UPC – R ecommended to evaluate forproteinuria • CBC with blood film evaluation – R ecommended as part of aminimum databaseHeartworm diseaseDirofilaria immitis, the causative agent of heartworm disease, is transmitted when mosquitoes infected with D. immitis larvae feed on (or bite) a healthy dog. Heartworm disease has subtle or mild clinical signs in the early stages, making preventive measures so much more important—especially as advanced infection may result in death.Did you know?•D espite availability of monthly preventives, prevalence rates of canine heartworm have remained consistent nationwide.7•T he American Heartworm Society (AHS) and the Companion Animal Parasite Council (CAPC) recommend testing all dogs for both antigen and microfilariae annually.7,8•F or more information and current recommendations on treating canine heartworm disease, goto or . Dirofilaria immitisPrimary vectorMosquitoPathologyInfective larvae (L3) mature to adult worms in the heart and pulmonary arteriesClinical presentation Asymptomatic at first, later developing:• Mild, persistent cough• Lethargy• Exercise intolerance• Reduced appetite• Weight lossLaboratory abnormalities that may be seen • Eosinophilia• Azotemia• Increased liver enzymes• ProteinuriaAnaplasmaphagocytophilumAnaplasma platysPrimary vectorsIxodes scapularis Ixodes pacificusBlack-legged tick (deer tick)Rhipicephalus sanguineus (brown dog tick)PathologyInfects neutrophilsInfects plateletsClinical presentationCan present acutely:• F ever • Anorexia • Lethargy• Polyarthritis, lameness • Neurologic signsUsually minimal clinical signs, but some dogs may have:• F ever • Uveitis• Petechiae and ecchymoses • EpistaxisLaboratory abnormalities• Thrombocytopenia• Anemia• Lymphopenia• Increased liver enzymesOther findings may be seen:• Decreased albumin • Increased globulin• Increased ALP and ALT • Proteinuria• Decreased Urine SG • Increased UPC NotePrevious infection may not prevent reinfection and persistent infections are possible.9,10Canine anaplasmosisCanine granulocytic anaplasmosis is caused by the bacterium Anaplasma phagocytophilum (transmitted by the black-legged tick [deer tick]). Anaplasma platys (transmitted by the brown dog tick) is the cause of infectious cyclic thrombocytopenia.Did you know?•M any mammalian species, including humans, are susceptible to A. phagocytophilum infection. •D ogs coinfected with Anaplasma and other bacterial pathogens may have more complex disease presentations and respond more slowly to therapy.• A .platys infects canine platelets and is frequently seen as a coinfection with Ehrlichia canis .Canine ehrlichiosisCanine ehrlichiosis is caused by the bacteria Ehrlichia canis (transmitted by the brown dog tick) and Ehrlichia ewingii (transmitted by the lone star tick). Canine Ehrlichia infections may progress to the subclinical phase or may become chronic infections.Ehrlichia canisEhrlichia ewingiiPrimary vectorRhipicephalus sanguineus (Brown dog tick)Amblyomma americanum (Lone star tick)PathologyInfects monocytes Infects granulocytesClinical presentation• Fever, anorexia, lethargy • Bleeding disorders • Polyarthritis, lameness • Lymphadenomegaly • Neurologic signs• Fever, anorexia, lethargy • Polyarthritis, lameness • Neurologic signsLaboratory abnormalitiesNotePrevious infection may not prevent reinfection, and persistent infections are possible.12,14Did you know?• D ogs coinfected with E. canis and A. platys were found to have more severe anemia and thrombocytopenia than dogs with either single infection.11• I n a study of healthy dogs with antibodies to E. canis , 39% were thrombocytopenic.12• C hronic E. canis infections, if left untreated, can lead to bone marrow dysfunction or kidney disease.• D ogs with Ehrlichia antibodies in E. canis endemic areas had a 112% increased risk of developing chronic kidney disease (CKD).13CKD monitoring• Chemistry panel with SDMA - R ecommended to evaluate forsecondary kidney disease.• Urinalysis with UPC - R ecommended to evaluatefor proteinuria • CBC with blood film - R ecommended as part of aminimum database• Anemia• Thrombocytopenia • Hyperglobulinemia • ProteinuriaOther clinical findings may include:• Decreased albumin • Increased globulin• Mild increased ALT and ALP • Increased SDMA • Creatinine• Decreased urine specific gravity, proteinuria • Increased urine protein:creatinine (UPC) ratio.Serology and PCR for sick patients For dogs presenting with clinical signs consistent with a vector-borne disease, usingserology and PCR together improves your ability to make an accurate diagnosis.Serology Polymerase chain reaction (PCR)Measures Antibody response of host Nucleic acid (DNA) from pathogenBenefits Useful for screening as well as diagnosisof infection Specifically identifies pathogens indicating active infectionLimitations Clinical signs may precede a measurableantibody response A negative PCR result does not necessarily rule out infectionDogs with ehrlichiosis and anaplasmosis may present with clinical signs at different times after infection. Which sick dog are you dealing with? Benefits and limitations of each diagnostic methodrecrudescence presents presents presentsWhen to use the IDEXX vector-borne disease RealPCR™ panels• S ick patients with clinical signsand/or laboratory abnormalitiesconsistent with a vector-borne illness• P atients with subclinical infectionsbased on history, physicalexamination, serology, and clinicallaboratory findings“No single test is sufficientfor diagnosing an infectious disease in a sick patient.”Edward Breitschwerdt, DVM, DACVIM*Professor, Internal MedicineCollege of Veterinary Medicine,North Carolina State University* D r. Breitschwerdt has a business relationship with IDEXX pursuant to which he receives compensation from IDEXX from time to time. The views expressed in this guide are solely those of Dr. Breitschwerdt.References1. G eneral guidelines: Parasite testing and protection guided by veterinarians [dog].Companion Animal Parasite Council website. /guidelines /general-guidelines. Updated July 29, 2020. Accessed November 17. 2021. 2. B eall MJ, Chandrashekar R, Eberts MD, et al. Serological and molecular prevalenceof Borrelia burgdorferi , Anaplasma phagocytophilum , and Ehrlichia species in dogs from Minnesota. Vector-Borne Zoonotic Dis . 2008;8(4):455–464. doi:10.1089/vbz.2007.0236 3. R egions where ticks live [maps]. Centers for Disease Control and Prevention website./ticks/geographic_distribution.html. November 17, 2021. 4. D rake C, Coyne M, McCrann DJ, Buch J, Mack R. Risk of development of chronickidney disease after exposure to Borrelia burgdorferi and Anaplasma spp. T op Companion Anim Med. 2020;42:100491. doi:10.1016/j.tcam.2020.100491 5. O ’Connor TP , Esty KJ, Hanscom JL, Shields P , Philipp MT . Dogs vaccinatedwith common Lyme disease vaccines do not respond to IR 6, the conserved immunodominant region of the VlsE surface protein of Borrelia burgdorferi.Clin Diagn Lab Immunol. 2004;11(3):458–462. doi:10.1128/CDLI.11.3.458-462.2004 6. S traubinger RK. PCR-based quantification of Borrelia burgdorferi organisms in caninetissues over a 500-day postinfection period. J Clin Microbiol. 2000;38(6):2191–2199. doi:10.1128/JCM.38.6.2191-2199.2000 7. C APC prevalence maps: heartworm [dog]. Companion Animal Parasite Councilwebsite. /maps/#/2021/all-year/heartworm-canine/dog/united-states. Accessed November 17, 2021.8. A merican Heartworm Society. Current canine guidelines for the prevention, diagnosis,and management of heartworm infection in dogs . 2020. Accessed November 17, 2021. https:///images/pdf/AHS_Canine_Guidelines_11_ 13_20.pdf?1605556516 9. E genvall A, Lilliehöök I, Bjöersdorff A, et al. Detection of granulocytic Ehrlichia speciesDNA by PCR in persistently infected dogs. Vet Rec . 2000;146(7):186–190. doi:10.1136/vr.146.7.186 10. B reitschwerdt EB, Hegarty BC, Qurollo BA, et al. Intravascular persistence ofAnaplasma platys , Ehrlichia chaffeensis , and Ehrlichia ewingii DNA in the blood of a dog and two family members. Parasit Vectors . 2014;7:298. doi:10.1186/1756-3305-7-298 11. G aunt S, Beall M, Stillman B, et al. Experimental infection and co-infection of dogs withAnaplasma platys and Ehrlichia canis : hematologic, serologic and molecular findings. Parasit Vectors . 2010;3(1):33. doi:10.1186/1756-3305-3-33 12. H egarty BC, de Paiva Diniz PP , Bradley JM, Lorentzen L, Breitschwerdt E. Clinicalrelevance of annual screening using a commercial enzyme-linked immunosorbentassay (SNAP 3Dx) for canine ehrlichiosis. J Am Anim Hosp Assoc. 2009;45(3):118–124. doi:10.5326/0450118 13. B urton W, Drake C, Ogeer J, et al. Association between exposure to Ehrlichia spp.and risk of developing chronic kidney disease in dogs. J Am Anim Hosp Assoc . 2020;56(3):159–164. doi:10.5326/JAAHA-MS-7012 14. S tarkey LA, Barrett AW, Beall MJ, et al. Persistent Ehrlichia ewingii infection indogs after natural tick infestation. J Vet Intern Med . 2015;29(2):552–555. doi:10.1111/jvim.12567 15. S nellgrove AN, Krapiunaya I, Ford SL, et al. Vector competence of Rhipicephalussanguineus sensu stricto for Anaplasma platys . Ticks Tick Borne Dis. 2020;11(6):101517. doi:10.1016/j.ttbdis.2020.101517Depend on the most accurate and comprehensive screenSNAP 4Dx Plus T estReference-laboratory quality in the palm of your hand, for superior diagnostic accuracy at the pointof care.Lab 4Dx Plus T estAvailable from IDEXX Reference Laboratories as a stand-alone test or as part of a more comprehensiveparasite screen with the Fecal Dx Antigen Panel with Lab 4Dx Plus Test-Canine.SNAP ® technology uses a proprietary three-step process to deliver dependable sensitivity and specificity.Available in-clinic or from IDEXX Reference LaboratoriesStrengthen the bonds.IDEXX Laboratories, Inc.One IDEXX DriveWestbrook, Maine 04092United StatesAmerican dog tick (Dermacentor variabilis) photographer: Susan E. Ellis, USDA-APHIS-PPQ. Black-legged tick (Ixodes scapularis), lone star tick (Amblyomma americanum), and brown dog tick (Rhipicephalus sanguineus) photographer: James L. Occi.© 2022 IDEXX Laboratories, Inc. All rights reserved. • 09-69074-13All ®/TM marks are owned by IDEXX Laboratories, Inc. or its affiliates in the United States and/or other countries. The IDEXX Privacy Policy is available at .。

人附睾蛋白4(HE4)对子宫内膜癌和卵巢癌的早期诊断价值

人附睾蛋白4(HE4)对子宫内膜癌和卵巢癌的早期诊断价值【摘要】目的比较血清人附睾分泌蛋白4( HE4)对于卵巢癌和子宫内膜癌患者的诊断价值。

方法实验分为5组:卵巢癌患者、卵巢良性疾病患者、子宫内膜癌患者、子宫内膜良性疾病患者和健康对照者，分别采用ELISA方法和化学发光方法对血清中的H E4水平和CA125水平进行检测。

结果卵巢癌及子宫内膜癌患者血清中HE4和CA125的含量均高于卵巢良性疾病组、子宫良性疾病组和健康对照组( P 0.05）；卵巢癌患者HE4的敏感度及特异度与CA125基本一致（p>0.05）。

结论对于子宫内膜癌患者和卵巢癌患者而言，HE4均具有较高的敏感性和特异性，对于疾病的早期诊断具有很大的辅助作用。

【关键词】卵巢癌，子宫内膜癌，HE4，诊断价值子宫内膜癌和卵巢癌是发生于女性生殖器官的肿瘤之一，两者的发病率均较高，目前对于两种疾病的诊断手段在不断发展，免疫学技术的应用也较为普遍，肿瘤标志物CA125对于诊断卵巢癌患者而言应用较为成熟，但是敏感性较低[1]；对于子宫内膜癌而言，目前临床上尚没有发现特异性的肿瘤标志物；人附睾蛋白4（HE4）是近年来研究较多的一个肿瘤标志物，因此，本文主要研究HE4 对于卵巢癌和子宫内膜癌患者进行早期诊断的价值。

1 资料与方法1.1一般资料本次研究共分为五组：（1）卵巢癌患者组（35例）：年龄在28~ 66岁之间，平均年龄51.7岁，其中有19例为浆液性腺癌、有10例为黏液性腺癌、有6例为低分化腺癌。

（2）卵巢良性患者组（36例）：年龄26~ 64 岁，平均50.9岁，其中有21例为子宫肌瘤、有15例为卵巢囊肿。

（3）子宫内膜癌患者组（35例）：年龄23~ 63 岁，平均50.4岁，均为早期患者；（4）子宫良性疾病患者组（37例）：年龄24～65 岁，平均49.8岁，其中有7例为卵巢囊肿患者，有8例为浆液性囊腺瘤患者，有16例为囊性畸胎瘤患者，有6例为黏液性囊腺瘤患者；（5）健康对照组（35例）：年龄24~ 62 岁, 平均50.6岁，所有患者均排除了心、肝、肺的不良疾病，同时多有受试者均无卵巢疾病史。

Euforea Pocket Guide for Allergic Rhinitis说明书

Pocket guide Allergic rhinitisDEVELOPED BY EUFOREA EXPERT TEAMS1What should the physician do?2Ask about allergic symptoms and the medical history ofyour patientDetermine the severity of the disease and impact of themajor symptomPerform physical examination, including anteriorrhinoscopyConfirm allergy by skin prick test or serum specific IgECheck for lower airway symptoms, especially asthmaWhen to suspect comorbid asthma?3Questions to your patientHave you had an episode or recurrent episodes ofwheezing?Do you have troublesome cough, especially at night/during awakening/excercise?Do you cough or wheeze after exercise?Do you experience extended common cold/laryngitis/bronchitis?Does your chest feel tight or do you feel impairedbreathing out?If YES to any of these question: your patient might beasthmatic.23(1) Greiner AN, et al. Lancet, 2011; 378:2112-22.(2) Adapted from: Scadding GK, et al. Clin Exp Allergy, 2017;47:856-889.(3) Adapted from: Bousquet J, et al. Allergy, 2008. 63 Suppl 86:8-160.(4) Caimmi D, et al. Clin Exp Allergy, 2017; 47:1526-1533.How to use the AR pocket guide in 5 steps1. Diagnose AR • History• Symptom(s)• Confirm sensitization 2. Classify patient• Symptom(s)• Treatment response2’. R eclassifypatientUncontrolled AR(VAS ≥ 5)Personalized treatment based on: • treatment response • long-term plan • patient needs 3. Define th • Medical trea • Patient educ • Patient part3’. R edefintherapy4Controlled AR (VAS < 5)6. Patient follow up4. Select product• Pharmacotherapy• Allergen Immunotherapy4’. S elect product5. A ctivate treatment plan5’. F inetune treatment plan5Uncontrolled7(*) Add-on therapies• Isolated watery rhinorrhoea: Ipratropium • R hinorrhoea in asthmatics: Leukotriene receptor antagonist • O cular itch/skin rash: Oral non-sedating anti-H1• O cular symptoms: Intra-ocular anti-H1 or Cromones • S udden onset nasal blockage: nasal / oral decongestant ≤ 7 daysRe-evaluate diagnosisAllergen Immunotherapy (moderate-severe AR)and/orOral corticosteroid (short course)and/or Surgery(severe nasal obstruction) and/or Add-on therapies (*)Allergen Immunotherapy(AR due to i.e. pollen or house dust mite)VAS ≥ 5(#) Depending on availability at national levelWhat is AIT?6AIT (also called desensitization, hyposensitization or allergy vaccination) is a treatment with administration of increasing amounts of anallergen to induce immunological tolerance and to prevent allergicsymptoms upon re-exposure. AIT can be administered via differentroutes: subcutaneous immunotherapy (SCIT), with s.c. injections of thesensitizing allergens in the upper arm, and sublingual immunotherapy(SLIT), with the sensitizing allergen kept under the tongue for 1-2 min(in the form of tablets or drops).What are the advantages of AIT?6Efficacy varies between specific productsO nly treatment with disease modifying capacityR educes nasal and/or ocular symptomsE nhances the quality of lifeL owers need for intake of other anti-allergic medicationI nduces immunological tolerance, providing sustained clinical benefitH as the potential to prevent asthma89Which patients can benefit from AIT?5AIT should be considered if ALL are present:Uncontrolled moderate-to-severe symptoms of AR +/- conjunctivitis, on exposure to clinically relevant allergensConfirmation of IgE sensitation to clinically relevant allergens (via skin prick test or serum specific IgE) Inadequate control of symptoms despite reliever medication and allergen avoidance measures and/or unacceptable adverse effects of medication(5) Roberts G, et al. Allergy, 2018; 73: 765-798.(6) Hellings PW, et al. Clin Transl Allergy, 2019; 9:1-7.10(5) Table adapted from: Roberts G, et al. Allergy, 2018; 73: 765-798. (*) Always adhere to product-specific SmPCAbsolute contra-indications for AIT5Always adhere to product-specific SmPCUncontrolled or severe asthmaActive, systemic auto-immune disorders, or other severediseaseActive malignant neoplasiaInitiation of AIT during pregnancyUnder the age of 5For relative contra-indications: contact specialist.11(5) Roberts G, et al. Allergy, 2018; 73: 765-798.。

人凝血酶原iv复合物

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文档下载后可定制随意修改，请根据实际需要进行相应的调整和使用，谢谢!并且，本店铺为大家提供各种各样类型的实用资料，如教育随笔、日记赏析、句子摘抄、古诗大全、经典美文、话题作文、工作总结、词语解析、文案摘录、其他资料等等，如想了解不同资料格式和写法，敬请关注!Download tips: This document is carefully compiled by the editor. I hope that after you download them, they can help yousolve practical problems. The document can be customized and modified after downloading, please adjust and use it according to actual needs, thank you!In addition, our shop provides you with various types of practical materials, such as educational essays, diary appreciation, sentence excerpts, ancient poems, classic articles, topic composition, work summary, word parsing, copy excerpts,other materials and so on, want to know different data formats and writing methods, please pay attention!人凝血酶原IV复合物，在医学领域中扮演着重要的角色。

程序性细胞死亡因子4是替代活化巨噬细胞的新分子标志物

程序性细胞死亡因子4是替代活化巨噬细胞的新分子标志物田稼;蒋晓刚;李海燕;钟波;张富军;宁启兰;韩燕;杨旭东【期刊名称】《中国组织工程研究》【年(卷),期】2015(019)051【摘要】背景：程序性细胞死亡因子4的表达水平与经典活化的巨噬细胞的表型负相关,但是程序性细胞死亡因子4的表达水平与巨噬细胞替代活化的关系仍不明确。

目的：观察替代活化的巨噬细胞中程序性细胞死亡因子4的表达水平改变,及其对替代活化的影响。

方法：用白细胞介素4、地塞米松单独刺激或联合刺激诱导NR 8383细胞的替代活化;采用qP CR检测巨噬细胞替代活化的分子标志物的表达水平,确定诱导巨噬细胞替代活化的最适条件;采用qP CR和Western blot检测替代活化的巨噬细胞模型中程序性细胞死亡因子4表达的改变;分别用大鼠程序性细胞死亡因子4高表达质粒和程序性细胞死亡因子4干扰质粒转染NR 8383细胞株,采用荧光倒置显微镜观察转染的效率,并检测程序性细胞死亡因子4的表达水平;分别检测程序性细胞死亡因子4高表达和程序性细胞死亡因子4敲低后NR 8383细胞株中巨噬细胞经典活化和替代活化的分子标志。

结果与结论：（1）白细胞介素4和地塞米松联合刺激比白细胞介素4或地塞米松单独刺激可更有效的诱导巨噬细胞的替代活化;10μg/L白细胞介素4＋50 nmol/L地塞米松刺激24 h可有效诱导巨噬细胞的替代活化。

（2）在替代活化的巨噬细胞中程序性细胞死亡因子4的表达水平显著升高,是替代活化的分子标志。

（3）程序性细胞死亡因子4高表达上调替代活化的分子标志的表达（P〈0.05）;而敲低程序性细胞死亡因子4可下调CD206的表达（P〈0.05）,并显著上调经典活化的分子标志诱导型一氧化氮合酶的表达（P〈0.05）。

结果表明程序性细胞死亡因子4上调是替代活化巨噬细胞的一个重要分子标志物。

【总页数】8页(P8281-8288)【作者】田稼;蒋晓刚;李海燕;钟波;张富军;宁启兰;韩燕;杨旭东【作者单位】[1]陕西省微生物研究所,陕西省西安市710043;[2]西安交通大学医学部基础医学院生物化学与分子生物学系,陕西省西安市710061;[3]西安交通大学医学部基础医学院药理学系,陕西省西安市710061;[4]西安交通大学第二附属医院儿科,陕西省西安市710004【正文语种】中文【中图分类】R318【相关文献】1.经典活化型/替代活化型巨噬细胞对血管平滑肌细胞凋亡的影响 [J], 尚茹茹;刘晓红;张锦;吕淑萍;李飞虹2.程序性细胞死亡因子4是替代活化巨噬细胞的新分子标志物 [J], 田稼;蒋晓刚;李海燕;钟波;张富军;宁启兰;韩燕;杨旭东3.积雪草酸对糖尿病肾病大鼠肾功能及巨噬细胞表面活化标志物水平的影响 [J], 罗先荣;彭家清;熊燕;钟广芝4.积雪草酸对糖尿病肾病大鼠肾功能及巨噬细胞表面活化标志物水平的影响 [J], 罗先荣;彭家清;熊燕;钟广芝;5.替代活化M2型巨噬细胞在增生性瘢痕形成中的作用研究 [J], 朱镇森因版权原因，仅展示原文概要，查看原文内容请购买。

人附睾蛋白4检测早期卵巢癌

人附睾蛋白4检测早期卵巢癌谷维【期刊名称】《国际妇产科学杂志》【年(卷),期】2015(42)2【摘要】卵巢癌是威胁女性生命健康的三大生殖道恶性肿瘤之一,其年轻化趋势明显且死亡率高居首位,如何提高卵巢癌患者早期检出率是降低该病死亡率的关键.传统卵巢癌标记物——糖类抗原125(CA125)虽然广泛应用于临床,但是敏感度和特异度较低,并且在一些妇科良性疾病中也出现非特异性的升高,因此迫切需要一种高敏感度和特异度的检测指标.近年来颇受瞩目的血清标记物——人附睾蛋白4(HE4)成为了早期诊断卵巢癌的焦点,相比于CA125,HE4在大多数非卵巢恶性肿瘤中不表达或低表达,并且单独检测HE4或联合CA125在对卵巢癌的早期诊断、疗效和预后评价等方面更具优势,现已在国外和国内多家三甲医院推广应用.现对HE4的发现、发展和临床应用及其与CA125构建的卵巢恶性肿瘤发病风险模型(ROMA)作综述.【总页数】4页(P141-144)【作者】谷维【作者单位】201508 上海,复旦大学附属金山医院妇产科【正文语种】中文【相关文献】1.血清人附睾蛋白4和糖类抗原125检测在早期卵巢癌诊断中的应用 [J], 江涛;周芳芳;李军;王昌富2.人附睾蛋白4及 CA125联合检测在卵巢肿瘤中的应用及人附睾蛋白4水平与卵巢肿瘤病理学分型的相关性研究 [J], 白杰3.CA125、HE4、ROMA值联合检测对早期卵巢癌风险的预测价值 [J], 舒琪;王丽萍;岳丽娟4.血清HE4、CA 125、SMRP及外周血TAP联合检测对早期卵巢癌的筛查价值[J], 张新新5.HE4、DJ-1、SMRP联合检测在早期卵巢癌诊断中的应用价值 [J], 李晓莹;齐红燕;叶敏因版权原因，仅展示原文概要，查看原文内容请购买。

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Figure How a Distributor Effects Economy of Effort
Case 1
迪比特忍着剧痛进行了全面的渠道革命。前天，迪比特执行副总裁彭新淼宣布，为了保住销售业绩，公司已经取消区域代理商，并用近5亿元的现金代价从经销商中购回几十万台手机，改由各省分公司直供各经销商。但迪比特并不是手机厂商渠道变革第一个，早在去年，诺基亚就对其渠道实施改革，并借此坐上了中国手机的“老大”位置。而摩托罗拉、康佳、NEC等手机厂商也不甘落后，纷纷对销售渠道实施“手术”。这一切都发生在手机市场将面临着重新洗牌的大背景下，谁也不愿意渠道拖后腿，而往往渠道就是决定手机厂商未来生存的一个极为重要的因素
4Channel factors影响分销渠道的因素
案例美国佩珀饮料公司在80年代初就成为非可乐类饮料世界排名第一的公司，在整个饮料行业里排名第三。佩珀公司把浓缩饮料卖给分装厂商，分装厂商将饮料稀释后装瓶，辅以广告促销，推销给零售商，再由零售商卖给消费者。佩珀公司与全美500多家分装厂商建立密切的关系，分装厂商的销售人员与零售商保持密切联系，制作本地的促销广告，要求零售商把佩珀饮料放再最显眼的柜台处，经常运用折扣、特殊陈列品、优惠券和免费样品来促进产品的销售。分装厂商十分了解当地的市场情况和零售商的需要，帮助佩珀公司制订各地区的营销方案。尽管这些分装厂商同时也经销可口可乐和百事可乐，但绝大多数都把佩珀饮料作为最佳品牌的饮料。
分销渠道的类型 types of marketing channels
所谓销售渠道抑或分销渠道皆是指：产品从生产者到最终消费者的途径。 A set of interdependent organizations involved in the process of making a product or service available for use or consumption by the consumer or business user. 1、销售渠道的层次 1.1直销 direct channels 没有中间层次的销售 1.2间接销售 indirect channels 中间机构包括：批发商、中转商和零售商。 Producers lose more control and face greater channel complexity as additional channel levels are added.
Chapter 8 place of channels
第八章分销渠道
Objectives:
5 Identify
value networks and marketingchannel systems. 5 Learn the type of work performed by marketing channels. 5 Understand the decisions companies face in designing, managing, evaluating, and modifying channels
Types of Vertical Marketing Systems
•
Corporate VMS Contractual VMS Administered VMS
•
•
3.2渠道冲突的管理Managing channel conflict
纵向营销系统 vertical marketing systems 通过合同或者其他方式将制造商、中间商和零售商联合起来成为整体的方法。其目的是解决渠道的纵向冲突。
Channel-Design Decisions
Channel Factors
3.1渠道组织与纵向营销系统 Vertical Marketing System
•
Hale Waihona Puke •A distribution channel structure in which producers, wholesalers, and retailers act as a unified system One channel member owns the other, has contracts with them, or has so much power that they all cooperate.
• Exclusive distribution
• Severely limited distribution
• Selective distribution
• Some intermediaries willing to carry good are selected
• Intensive distribution
2渠道的作用Work Performed by Channels
Offers the firm more than it can achieve on its own through the intermediaries: •Contacts •Experience •Specialization •Scale of operation
3渠道行为和渠道冲突Channel Behavior and channel conflicts
• The channel will be most effective when: • each member is assigned tasks it can do best. • all members cooperate to attain overall channel goals. • If this does not happen, conflict occurs: • Horizontal Conflict occurs among firms at the same level of the channel (e.g., retailer to retailer). • Vertical Conflict occurs between different levels of the same channel (e.g., wholesaler to retailer). • Some conflict can be healthy competition.
Case 2
2004年下半年，国内影碟机市场巨头夏新在经过反复考证衡量之后，对影碟机的销售渠道进行了主动变革，全面接盘广东一家以“苹果熟了” 而红及一时的某碟机企业的渠道精英，补充和整合夏新影碟机在部分区域市场的销售渠道。
Channel Functions
• • • •
Information Promotion Contact Matching
4Channel-Design Decisions
•
• • •
Channel Factors
Intermediary type Number of intermediaries Terms and responsibilities of intermediaries cost
• Company sales force • Manufacturer’s agency • Industrial distributors
•
Channel-Design Decisions
Independent insurance agents act for multiple insurance providers
Channel-Design Decisions
•
• • •
Channel Factors
Intermediary type Number of intermediaries Terms and responsibilities of intermediaries cost
4Channel factors影响分销渠道的因素
1982年，佩珀公司改变依靠分装厂商在当地做广告促销的方法，实行全国统一的集中营销方案。公司削减了地方性的销售人员，减少了对分装厂商的业务支持，以全国性的广告活动取代了过去由分装厂商在当地做促销广告的做法。当年公司销售量下降了3%，到秋季时该公司亏损4000万美元，市场排名下降至第四名。分装厂商与公司的关系也日益疏远，开始对公司采取防备状态。在这一事例中，佩珀公司在分销渠道选择时出现了什麽失误？你认为在选择分销渠道是应该注意哪些因素？
Pricing, Marketing communications
A strong distribution system can be a competitive advantage Channel decisions involve long-term commitments to other firms
Objectives:
Identify trends taking place in channel dynamics. 5 Learn how channel conflict can be managed.
5
The importance of marketing channels
Channel choices affect other decisions in the marketing mix
• Offering is placed in as many outlets as possible.
•
Number of Intermediaries
中间商特性（不一定非要拥有零售渠道的所有权，会影响其他代理渠道的积极性，通过其他形式也可以加强控制）如何使分销商忠诚：奖励性佣金，会增加分销成本。企业品牌忠诚度。培训机会和奖励旅游。
零售商合作社
特许专卖机构
3.3Horizontal marketing system平行营销系
统