乳腺癌患者术后预后生存质量的相关因素及中性粒细胞/淋巴细胞比值Ki67指数对预后生存质量不良的预测价值

目的 探讨乳腺癌患者术后预后生存质量的相关因素及中性粒细胞/淋巴细胞比值(NLR)、Ki67指数对预后生存质量不良的预测价值。方法 选取2021年1月—2021年12月丽水市人民医院收治的90例乳腺癌患者为研究对象，其中采用传统根治术42例，改良根治术48例，统计所有乳腺癌患者术后预后生存质量，分析预后生存质量的相关因素及对NLR、Ki67指数对预后生存质量不良的预测价值。结果 90例乳腺癌患者，术后预后良好69例(76.67%),预后不良21例(23.33%)。单因素分析所示，乳腺癌患者术后预后生存质量与年龄、绝经状态、体质量及乳腺癌家族史无关(P>0.05);乳腺癌患者术后预后生存质量与病理类型、肿瘤大小、肿瘤分期、脉管瘤栓、腋窝淋巴结状态、术后是否接受化疗、手术方式、皮下积液是否发生、NLR及Ki67指数有关，与预后良好患者相比，预后不良患者浸润性、肿瘤≥5 cm、Ⅲ～Ⅳ期、脉管瘤栓阳性、腋窝淋巴结状态阳性、术后未接受化疗、传统根治术、发生皮下积液、NLR≥10×109/L及Ki67指数≥40%构成比较高(P<0.05)。多因素logistic回归分析，结果显示病理类型、肿瘤大小、肿瘤分期、脉管瘤栓、腋窝淋巴结状态、术后是否接受化疗、手术方式、皮下积液是否发生、NLR及Ki67指数为影响乳腺癌患者预后生存质量的主要因素(P<0.05)。两项联合乳腺癌术后预后生存质量不良的预测灵敏度、准确性高于NLR、Ki67指数单一检测(P<0.05)。结论 乳腺癌患者术后预后生存质量与病理类型、肿瘤大小、肿瘤分期、脉管瘤栓、腋窝淋巴结状态、术后是否接受化疗、手术方式、皮下积液是否发生、NLR及Ki67指数相关，NLR、Ki67指数联合检测对预后生存质量不良的预测价值较高。     

PRL-3对三阴性乳腺癌患者肝转移的预测意义及其与生存关系的研究

目的探讨PRL-3在三阴性乳腺癌(TNBC)组织中的表达情况,以及不同的表达水平对患者肝脏转移的预测意义及其与生存的关系。方法收集三阴性乳腺癌患者术后组织标本及临床资料,采用免疫组化法和Western Blot法检测组织中PRL-3不同的表达水平,分析不同表达水平与临床病理特征的关系及与肝转移的相关性,采用Kaplan-Meier生存分析其预后。结果三阴性乳腺癌组织中PRL-3的检测阳性率为67.47%,阳性表达与乳腺癌患者年龄、核分级、P53、肿瘤大小无相关性,与TNM分期、淋巴结状况存在相关性,临床分期越晚其表达量越高。PRL-3阳性组的肝转移发生率明显高于阴性组,差异有统计学意义。PRL-3阴性与阳性组中位DFS分别为:24.7、16.5个月,中位OS分别为41.6、30.8个月,差异均有统计学意义。结论 PRL-3在三阴性乳腺癌中过度表达,促进癌细胞的侵袭,与肝脏转移密切相关,可作为评估TNBC预后的潜在生物标志物。     

中性粒细胞淋巴细胞比率及血小板淋巴细胞比率在乳腺癌合并感染患者术后生存期的预测研究

目的研究中性粒细胞淋巴细胞比率(NLR)及血小板淋巴细胞比率(PLR)在乳腺癌合并感染患者术后生存期的预测,为提高生存期提供依据。方法选取2010年10月-2015年10月医院诊治89例乳腺癌合并感染患者资料进行分析;采用自拟问卷调查表对患者基本资料进行统计,计算患者治疗前后PLR和NLR指标,分析两指标对乳腺癌合并感染患者术后生存期的影响。结果患者治疗后PLR≥300患者5年总生存率(Overall survival,OS)为72.8%;3年无病生存率(Disease-free survival,DFS)为30.8%;治疗前150PLR300患者5年OS为91.2%,3年DFS为69.4%;治疗后PLR150患者OS为100.0%;3年DFS为99.1%,患者治疗后不同PLR OS生存率和DFS生存率差异有统计学意义(P0.05);患者治疗后对于NLR≥5患者5年OS为76.1%;3年DFS为31.2%;治疗后对于NLR5组患者5年OS为98.3%,3年DFS为91.8%。患者治疗后不同NLR下OS及DFS生存率差异有统计学意义(P0.05)。结论 NLR及PLR能够准确的预测乳腺癌合并感染患者术后生存期,正确的指导患者临床治疗,值得推广应用。     

胃癌患者根治术后外周血中性粒细胞与淋巴细胞比值影响预后的研究

目的探讨根治术后胃癌患者血常规中中性粒细胞与淋巴细胞绝对值的比值(NLR)与预后的关系。方法选取有完整术后病理及随访资料的根治术后胃癌患者共215例,所有患者未接受术前新辅助放、化疗,计算每例患者术后的NLR,结合临床、病理参数指标,COX多因素回归分析各因素对术后无病生存(DFS)的影响,Kaplan-Meier方法绘制术后DFS曲线并计算中位DFS,比较不同NLR时DFS的差异。结果卡方分析显示,术后NLR水平与肿瘤的T分期相关。COX回归分析表明,T分期、肿瘤分化及NLR是影响术后DFS的独立预后因素(P<0.05)。术后NLR≤6.8与NLR>6.8的患者,术后中位DFS分别是69个月和56个月(P<0.001)。结论术后NLR水平是影响胃癌根治术后生存的独立预后因素。     

干细胞标志物ALDH1与大样本乳腺癌患者的预后关系

目的研究干细胞标志物已醛脱氢酶1(ALDH1)在乳腺癌细胞中的表达及其与乳腺癌患者预后的关系。方法采用免疫组化方法检测ALDH1在595例乳腺癌组织芯片中的表达并评分,用ROC方法确定ALDH1表达最佳临界值,检测ALDH1表达情况与患者临床特征之间相关性。结果 ALDH1在乳腺癌细胞中高表达,并与Ki67、CEA和分子分型呈正相关。单因素和多因素分析均表明ALDH1可以作为乳腺癌OS、DFS独立预后因子。结论 ALDH在乳腺癌组织中高表达,且过表达ALDH1可以作为乳腺癌独立预后不良因子。     

淋巴细胞/单核细胞比值在三阴性乳腺癌预后评估中的临床研究

目的 初步探讨外周血中淋巴细胞和单核细胞绝对值的比值（Lymphocyte to Monocyte Ratio,LMR）与三阴性乳腺癌预后的相关性,及与总生存时间的关系。方法 回顾性分析2010年11月1日至2015年12月31日辽宁省肿瘤医院110例行乳腺癌根治术三阴性乳腺癌患者临床病理资料,根据患者术前外周静脉血LMR分为低LMR组（LMR＜4.24,50例）和高LMR组（LMR≥4.24,60例）;采用Kaplan-Meier 生存分析法绘制生存曲线,比较两组患者的5年生存率,分析两组患者临床病理因素与总生存时间的关系。结果 LMR与三阴性乳腺癌之间具有相关性,低LMR组5年生存率为30.00%,高LMR组5年生存率为66.67%;两组相比较,5年生存率差异具有统计学意义（χ2=25.475,P＜0.05）;经Log Rank统计学分析,两组总生存时间相比较,差异具有统计学意义（χ2=6.932,P＜0.05）。结论 在三阴性乳腺癌患者中,术前外周血LMR可作为一个独立的危险因素,影响三阴性乳腺癌患者的预后。     

胃癌患者术后多项免疫分子表达对预后的影响

目的总结手术切除Ⅰ-Ⅲ期胃癌的临床特点,探讨胃癌术后常规检测分子表皮生长因子受体1(HER1)、HER2、P53、血管内皮生长因子(VEGF)、拓扑异构酶IIα(TOPIIα)、Ki67、P糖蛋白(P170)的表达及对预后的影响。方法回顾性分析41例I-III期胃癌手术病例,综合临床、病理特征及术后7项常规分子标记物的免疫组织化学指标,对患者无疾病生存时间(DFS)采用SPSS 17.0软件包进行单因素及多因素分析。结果全部病例术后随访时间为36个月,中位随访期18个月,1年总生存率为95.12%,中位DFS为17个月,1年DFS生存率为70.59%。术后患者HER1、HER2、P53、P170、Ki67、VEGF和TOPIIα均有阳性表达,HER2、P53、VEGF和TOPIIα高表达DFS降低(P0.05)。结论 HER2、P53、VEGF和TOPIIα阳性表达DFS降低。     

三阴性乳腺癌临床病理特点及与ki67表达关系的分析

目的:探讨三阴性乳腺癌的临床病理特点和预后特点以及ki67的表达情况与预后的关系。方法:选取浸润性乳腺癌患者271例,对其临床病理资料进行回顾性分析,统计随访结果和病理特征,并检测ki67的表达。结果:三阴性乳腺癌与非三阴性乳腺癌患者在绝经状态和肿瘤临床分期、淋巴结转移中ki67的表达存在差异且具有统计学意义(P<0.05)。相对于非三阴性患者,三阴性乳腺癌患者的生存率较低,差异有统计学意义(P<0.05)。ki67阳性的三阴性乳腺癌患者易出现淋巴结转移。结论:三阴性乳腺癌中Ki67阳性可作为患者不良预后的重要指标,对于指导临床治疗方案的制订具有重要意义。     

新辅助化疗对三阴性乳腺癌患者Ki67的影响

目的探讨新辅助化疗对三阴性乳腺癌(Triple Negative Breast Cancer,TNBC)患者核抗原Ki67的影响。方法选取2012年6月~2014年3月期间我院新辅助化疗治疗的TNBC患者136例,依据并采用免疫组化法检测所有患者乳腺癌组织中Ki67表达情况,化疗前分为阳性组和阴性组,化疗后依据Ki67表达改变情况阳转阴(A)组、阴转阳(B)组、阳不变(C)组和阴不变(D)组,随访5年,统计分析所有患者化疗疗效和生存情况。结果化疗前,阳性组患者化疗疗效明显优于阴性组,差异有统计学意义(p<0.05);化疗后,在疗效方面,A组>C组>D组>B组,差异有统计学意义(p<0.05);在5年内生存率方面,A组>C组>D组>B组,差异有统计学意义(p<0.05)。结论新辅助化疗可致使TNBC癌患者Ki67的表达发生转变,有可能改善患者生存预后,值得临床作进一步研究。     

年轻女性乳腺癌临床生物学特性及预后因素分析

目的 分析年轻女性(年龄≤35岁)乳腺癌患者临床生物学特性和预后,初步探讨年轻女性乳腺癌患者的预后因素、治疗策略及其相关性.方法 选取2003年1月至2008年4月经术后病理确诊的97例年轻女性乳腺癌患者,观察患者的生存与复发情况,探讨临床生物学特性与预后的相关性.结果 97例患者3年总生存率(OS)和3年无瘤生存率(DFS)分别为73.2％( 71/97)和64.9％(63/97).单因素分析显示临床分期、肿瘤大小、腋窝淋巴结转移、雌激素受体(ER)、手术方式、辅助化疗及内分泌治疗是年轻女性乳腺癌患者预后的相关因素(P＜0.01或＜0.05).多因素Cox回归分析显示,ER阴性较ER阳性患者的OS和DFS为差(RR=2.949,P=0.015;RR=2.629,P=0.007),未行内分泌治疗较行内分泌治疗患者的OS为差(RR=3.179,P=0.038).人类表皮生长因子受体2(HER-2)阳性较HER-2阴性患者的DFS为差(RR=2.079,P=0.032);临床分期Ⅱ、Ⅲ期较Ⅰ期患者的DFS为差(RR=2.801,P=0.017;RR=8.673,P=0.000);未行辅助化疗较行辅助化疗患者的DFS为差(RR=3.048,P=0.005).结论 ER阴性可作为独立的预后危险因素,HER-2阳性可作为年轻女性乳腺癌患者独立的疾病复发预测因素,术后行辅助化疗患者预后好,行内分泌治疗效果确切.     

中性粒细胞与淋巴细胞比率评估胃癌预后的Meta分析

目的分析探讨中性粒细胞与淋巴细胞比率(neutrophil-lymphocyte ratio, NLR)对胃癌患者预后评估的价值。 方法采用循证医学的meta分析研究。首先,从Embase、Medline和Web of Science数据库中检索关键词gastric cancer、stomach cancer、gastric carcinoma、stomach carcinoma、gastric neoplasm、stomach neoplasm、neutrophil和lymphocyte。然后,根据各纳入研究的总生存率(overall survival, OS)和无进展生存期(progression-free survival, PFS)/无疾病生存期(disease-free survival, DFS)合并生存危险比(hazard ratio, HR)的异质性,决定用固定或随机效应模型计算OS和PFS/DFS的合并HR。若合并HR>1,且其95%可信区间(confidence interval, CI)也>1, 则为NLR升高与胃癌患者OS或PFS/DFS的降低有显著联系。 结果收集18篇文献共纳入5 065例胃癌患者进行meta分析。升高的NLR与OS降低存在显著关联(HR为1.79,95%CI 1.54~2.08)。亚组分析中,胃癌多种治疗方案组和化疗组NLR升高均与OS降低存在显著关联[HR分别为1.84, (95% CI 1.48~2.29)和1.69, (95% CI 1.41~2.03)];而NLR≤3.2组和NLR>3.2组亦均与OS的降低存在显著关联[HR分别为1.80, (95% CI 1.46~2.23)和1.93, (95% CI 1.58~2.36)]。在胃癌患者OS的单因素meta回归分析中, 发表年份、种族、NLR临界值、治疗方案、患者数、进展期患者比例和男性比例都不是引起胃癌患者异质性来源的可能原因(P值分别为0.585、0.887、0.731、0.697、0.613、0.877、0.775)。 结论胃癌患者中升高的NLR与OS降低存在显著的关联,NLR可作为评价胃癌患者预后的标志物。     

The Prognostic Nutritional Index Predicts Survival and Identifies Aggressiveness of Gastric Cancer

Nutritional status has been associated with long-term outcomes in cancer patients. The prognostic nutritional index (PNI) is calculated by serum albumin concentration and absolute lymphocyte count, and it may be a surrogate biomarker for nutritional status and possibly predicts overall survival (OS) of gastric cancer. We evaluated the value of the PNI as a predictor for disease-free survival (DFS) in addition to OS in a cohort of 314 gastric cancer patients who underwent curative surgical resection. There were 77 patients in PNI-low group (PNI ≤ 47.3) and 237 patients in PNI-high group (PNI > 47.3). With a median follow-up of 36.5 mo, 5-yr DFS rates in PNI-low group and PNI-high group were 63.5% and 83.6% and 5-yr OS rates in PNI-low group and PNI-high group were 63.5% and 88.4%, respectively (DFS, P < 0.0001; OS, P < 0.0001). In the multivariate analysis, the only predictors for DFS were PNI, tumor-node-metastasis (TNM) stage, and perineural invasion, whereas the only predictors for OS were PNI, age, TNM stage, and perineural invasion. In addition, the PNI was independent of various inflammatory markers. In conclusion, the PNI is an independent prognostic factor for both DFS and OS, and provides additional prognostic information beyond pathologic parameters.     

Postoperative radiotherapy in patients with salivary duct carcinoma: clinical outcomes and prognostic factors

This study sought to investigate the clinical outcome and the role of postoperative radiotherapy for patients with salivary duct carcinoma (SDC) who had undergone surgery and postoperative radiotherapy. We performed a retrospective analysis of 25 SDC patients treated between 1998 and 2011 with surgery and postoperative radiotherapy. The median prescribed dose was 60 Gy (range, 49.5–61.4 Gy). The clinical target volume (CTV) was defined as the tumor bed in four patients, the tumor bed and ipsilateral neck in 14 patients, and the tumor bed and bilateral neck in six patients. Local control (LC), disease-free survival (DFS) and overall survival (OS) were estimated using the Kaplan-Meier method, and prognostic variables were analyzed with the log-rank test. The 5-year LC, DFS and OS were 67%, 45% and 47%, respectively. Disease recurrence was found in 12 patients: seven as local, four as regional and eight as distant failure. Perineural and lymphovascular invasion was a significant prognostic factor for LC (P = 0.03). Local failure was common, and the presence of local recurrence significantly affected the OS (P < 0.05). We conclude that surgery and postoperative radiotherapy is expected to decrease the risk of local failure and contribute to good prognoses for patients with SDC. It might be advisable to have the CTV include the cranial nerves involved and the corresponding parts of the skull base in cases of pathologically positive perineural invasion.     

外周血中性粒细胞与淋巴细胞比值评估小细胞肺癌患者预后的价值

目的 探讨治疗前外周血中性粒细胞与淋巴细胞比值（neutrophil-to-lymphocyte ratio,NLR）与小细胞肺癌患者预后的关系。方法 回顾性分析2013年1月~2015年3月安徽医科大学第一附属医院病理诊断明确的50例小细胞肺癌患者,所有患者未行手术治疗,以NLR中位数将患者分为高NLR组和低NLR组,对可能影响患者预后的各项因素进行分析。结果 47例病人获得随访。47例患者中1年生存率为53.2%。其中低NLR组的病人1年生存率为83.3%;高NLR组病人1年生存率为21.7%,单因素分析显示小细胞肺癌患者的预后与患者的分期、体力状况ECOG评分以及NLR水平相关（均有P<0.05）,对与预后相关的变量进行COX多因素分析,结果显示分期处于广泛期和高NLR水平是影响患者预后的独立危险因素（均有P<0.05）。结论 NLR可能是小细胞肺癌患者预后的独立危险因素。     

非典型脑膜瘤的临床特征及术后复发影响因素分析

目的探讨非典型脑膜瘤(AM)患者的临床特征及术后复发的影响因素。方法对2008年5月-2014年5月我院收治的48例AM患者的临床资料进行回顾性分析。结果本组AM患者发病大多50岁,且肿瘤部位主要位于大脑凸面,临床症状主要以头痛为首发表现,术前Simpson分级中Ⅰ~Ⅱ级较多见,免疫组化检查Ki-67指数5%~10%最常见。本组术后复发20例(41.67%),中位无复发生存期为4.6个月,中位生存期为12.96个月。复发者Simpson分级(Ⅲ~Ⅴ级)、Ki-67指数(30%)、AM形状(蘑菇形)及瘤周水肿程度(重度)比例高于无复发者(P0.05)。结论 AM具有术前诊断困难,高复发率及预后不良等临床特点,Simpson分级、Ki-67指数、AM形状及瘤周水肿程度为影响AM术后复发的独立危险因素。     

K-ras基因状况预测转移性结直肠癌药物治疗后生存期

目的旨在通过前瞻性检测转移性结直肠癌的K-ras基因表达状况,观察比较化疗单用或联合靶向治疗的长期疗效,为临床医生提供可靠预测指标。方法初治的转移性结直肠癌62例,一线化疗前全部检测了癌组织K-ras基因,62例癌组织K-ras基因进行检测,突变率为30.6%(19/62),其中G12占89.5%(17/19),G13占10.5%(2/19);43例K-ras野生型者分两组进行姑息化疗,包括单纯化疗组及化疗联合靶向治疗组,治疗期间密切随访,统计疗效及毒性并进行统计学分析。结果 K-ras突变者中位PFS和OS分别为4.7月和14.5月,野生型者中位PFS和OS分别为8.5月和24.0月,差异均有统计学意义(Log-rank检验P﹤0.05);10名K-ras野生型者在化疗基础上加用靶向治疗(西妥昔单抗),其中位生存期36.5月,显著高于未用靶向治疗患者的18.0月(Log-rank检验P﹤0.01)。结论癌组织K-ras基因突变者生存期明显短于K-ras基因野生型者,表明K-ras基因突变可作为本组转移性结直肠癌的有效预后预测指标;化疗联合靶向治疗转移性结直肠癌,可有效改善K-ras基因野生型者的PFS和OS,值得推广应用。     

Treatment outcomes of patients with FIGO Stage I/II uterine cervical cancer treated with definitive radiotherapy: a multi-institutional retrospective research study

The purpose of this study was to analyze the patterns of care and outcomes of patients with FIGO Stage I/II cervical cancer who underwent definitive radiotherapy (RT) at multiple Japanese institutions. The Japanese Radiation Oncology Study Group (JROSG) performed a questionnaire-based survey of their cervical cancer patients who were treated with definitive RT between January 2000 and December 2005. A total of 667 patients were entered in this study. Although half of the patients were considered suitable for definitive RT based on the clinical features of the tumor, about one-third of the patients were prescribed RT instead of surgery because of poor medical status. The RT schedule most frequently utilized was whole-pelvic field irradiation (WP) of 30 Gy/15 fractions followed by WP with midline block of 20 Gy/10 fractions, and high-dose-rate intracavitary brachytherapy (HDR-ICBT) of 24 Gy/4 fractions prescribed at point A. Chemotherapy was administered to 306 patients (46%). The most frequent regimen contained cisplatin (CDDP). The median follow-up time for all patients was 65 months (range, 2–135 months). The 5-year overall survival (OS), pelvic control (PC) and disease-free survival (DFS) rates for all patients were 78%, 90% and 69%, respectively. Tumor diameter and nodal status were significant prognostic indicators for OS, PC and DFS. Chemotherapy has potential for improving the OS and DFS of patients with bulky tumors, but not for non-bulky tumors. This study found that definitive RT for patients with Stage I/II cervical cancer achieved good survival outcomes.     

Neutrophil-lymphocyte ratio and nutritional status are clinically useful in predicting prognosis in colorectal cancer patients

Abstract

Background: Neutrophil-lymphocyte ratio (NLR) and nutritional status may provide a prognostic value in colorectal cancer (CRC). Thus, aim of this study was to evaluate the prognostic value of nutritional status and NLR in CRC patients.

Methods: A retrospective analysis was conducted in CRC patients. The independent variables were body mass index (BMI), weight loss (WL) and NLR. Logistic regression was used to estimate the odds chance of low NLR. Kaplan-Meier curves and Cox regression were used to evaluate the overall survival at 5?years old.

Results: In the 148 patients evaluated, the most prevalent nutritional status was overweight/obesity (43.2%) and 27.0% had severe WL. Sixty-seven subjects (45.3%) had NLR ≥ 3 that was associated with the lower OS (P?<?0.001). There was a higher OS for overweight/obese patients (P?=?0.002) and a lower among subjects with severe WL (P?=?0.009). The NLR ≥3 (HR: 3.639; 95% CI, 1.708–7.771) was an independent poor prognostic factor for OS. Patients without WL (HR: 0.367, 95% CI, 0.141–0.954) and classified as overweight/obesity (HR: 0.260; 95% CI, 0.106–0.639) presented better prognostic.

Conclusion: NLR, WL, BMI assessments are promising prognostic indicators in the CRC.     

Quality of life as a prognostic factor of overall survival in patients with advanced hepatocellular carcinoma: results from two French clinical trials

AIM: The aims of our study were to assess quality of life (QoL) as a prognostic factor of overall survival (OS) and to determine whether QoL data improved three prognostic classifications among French patients with advanced hepatocellular carcinoma (HCC). METHODS: We pooled two randomized clinical trials conducted by the Fédération Francophone de Cancérologie Digestive in a palliative setting. In each trial QoL was assessed at baseline using the Spitzer QoL Index (0-10). Three prognostic classifications were calculated: Okuda, Cancer of the Liver Italian Program (CLIP), and Barcelona Clinic Liver Cancer group (BCLC) scores. To explore whether the scores could be improved by including QoL, univariate Cox analyses of all potential baseline predictors were performed. A final multivariate Cox model was constructed including only significant multivariate baseline variables likely to result in improvement of each scoring system. In order to retain the best prognostic variable to add for each score, we compared Akaike information criterion, likelihood ratio, and Harrell's C-index. Cox analyses were stratified for each trial. RESULTS: Among 538 included patients, QoL at baseline was available for 489 patients (90%). Longer median OS was significantly associated with higher Spitzer scores at baseline, ranging from 2.17 months (Spitzer=3) to 8.93 months (Spitzer=10). Variables retained in the multivariate Cox model were: jaundice, hepatomegaly, hepatalgia, portal thrombosis, alphafetoprotein, bilirubin, albumin, small HCC, and Spitzer QoL Index (hazard ratio=0.84 95% CI [0.79-0.90]). According to Harrell's C-index, QoL was the best prognostic variable to add. CLIP plus the Spitzer QoL Index had the most discriminating value (C=0.71). CONCLUSIONS: Our results suggest that QoL is an independent prognostic factor for survival in HCC patients with mainly alcoholic cirrhosis. The prognostic value of CLIP score could be improved by adding Spitzer QOL Index scores.