济南市手足口病重症病例危险因素的病例对照研究

目的 探讨济南市手足口病重症病例发病的危险因素,为早期识别重症病例,降低手足口病重症病例的发生提供科学依据。 方法 采用病例对照研究方法,病例组为86例手足口病重症病例,对照组为86例手足口病轻症病例,用问卷调查收集病例组和对照组的社会人口学特征、接触史、就诊史、临床症状、实验室诊断等信息,对重症病例危险因素分析采用单因素和多因素非条件logistic回归分析。 结果 单因素结果显示发病到就诊日期、初诊单位类型为村级、患儿出现发热、足部皮疹、口腔疱疹、臀部皮疹、咳嗽、呕吐、有病例接触史、EV71阳性在两组间差异有统计学意义。多因素非条件Logistic回归分析结果显示,患儿出现发热(OR=89.38)、口腔疱疹(OR=22.28)、臀部皮疹(OR=234.81)、呕吐(OR=23.90)、有病例接触史(OR=31.48)和EV71阳性(OR=71.65)是手足口病重症病例发病的危险因素。 结论 早期识别发热、口腔疱疹、臀部皮疹、呕吐等症状的患儿,对降低手足口病重症病例的发生具有一定的参考意义。提高EV71手足口病疫苗的接种率,可降低由EV71导致的重症手足口病病例的发生率。     

A novel p.Gly417Valfs*12 mutation in the MTTP gene causing abetalipoproteinemia: Presentation of the first patient in Mexico and analysis of the previously reported cases

BackgroundOur aims were to describe the first Mexican patient with abetalipoproteinemia and to perform a comparative analysis of biochemical, clinical, and genetic characteristics of 100 cases reported in the literature.MethodsWe performed biochemical and molecular screenings in a Mexican girl with extremely low lipid levels and in her family. Further, we integrated and evaluated the characteristics of the cases with abetalipoproteinemia described in the literature.ResultsOur patient is a six‐year‐old girl who presented vomiting, chronic diarrhea, failure to thrive, malabsorption, acanthocytosis, anemia, transaminases elevation, and extremely low lipid levels. MTTP gene sequencing revealed homozygosity for a novel mutation p.Gly417Valfs*12 (G deletion c.1250). With the analysis of the reported cases, 60 clinical features (14 classical and 46 non‐classical) were observed, being the most common acanthocytosis (57.5%), malabsorption (43.7%), and diarrhea (42.5%); 48.8% of the patients presented only classic clinical features, while the remaining 51.2% developed secondary effects due to a fat‐soluble vitamin deficiency. An odds ratio analysis disclosed that patients diagnosed after 10 years of age have an increased risk for presenting clinical complications (OR = 18.0; 95% CI 6.0‐54.1, p < 0.0001). A great diversity of mutations in MTTP has been observed (n = 76, being the most common p.G865X and p.N139_E140) and some of them with possible residual activity.ConclusionThe first Mexican patient with abetalipoproteinemia presents a novel MTTP mutation p.Gly417Valfs*12. Three factors that could modulate the phenotype in abetalipoproteinemia were identified: age at diagnosis, treatment, and the causal mutation.     

Mandibular rescue: Application of the ALT fascia free flap to arrest osteoradionecrosis of the mandible

ObjectivesTo evaluate the use of the anterolateral thigh fascia free flap for use in neovascularization of mandibular bone in moderate osteoradionecrosis (ORN). All patients had ORN secondary to prior radiation therapy that was not severe enough to warrant segmental resection and reconstruction.Study designCase series.SettingTertiary medical center.MethodsIRB approval was obtained, and a retrospective chart review performed of all mandibular rescue procedures performed from 2011 to 2014. Patients with a minimum of two years of follow-up were included in the study.ResultsAll surgeries were performed by the senior surgeon (MF). Eight patients underwent the mandibular rescue procedure with resolution of pain and return to oral feeding in all patients, and no evidence of ORN progression on follow-up imaging. A total of 9 ALT free flaps were performed (one patient had 2 surgeries). Gender was distributed evenly (4 female/4 male). The average age was 66 (58-78), average length of hospitalization was 2.8 days (1–7), and average follow-up was 46.5 months (25–63).ConclusionsThe mandibular rescue procedure is a novel technique using the ALT fascia lata free flap to provide coverage and nutrient blood flow to mandible devascularized secondary to radiation therapy. The flap provides the advantages of low morbidity, ease of harvest, two-team approach to ablation and reconstruction, and quick recovery resulting in ‘short-stay’ free flap surgery. Although conclusions must be tempered in this small case series, our early clinical experience shows the ALT fascia lata flap holds promise in halting the destructive progression of ORN that is not yet advanced enough to require a segmental resection and reconstruction.     

吲哚菁绿造影在乳房重建中的应用进展

目的对吲哚菁绿（indocyanine green，ICG）造影在乳房重建中的应用进展进行综述。方法查阅国内外有关 ICG 造影在乳房重建中应用的文献，总结 ICG 造影历史沿革、化学成分、使用原理、使用方法及注意事项。结果ICG 是一种适用于术中血管灌注造影成像的荧光物质。ICG 造影及 SPY 灌注评估系统已逐渐用于乳房重建术中，包括组织扩张器及假体植入乳房重建和自体组织乳房重建，其与术者主观评估、手持多普勒血流探测仪、组织血氧测量及荧光素钠造影相比，有着明显优势及更多用途。结论ICG 造影可较准确评估乳房重建术中皮瓣灌注情况，从而指导术中决策，降低术后皮瓣相关并发症发生，并具有一定经济学效益。     

Development of a high-growth enterovirus 71 vaccine candidate inducing cross-reactive neutralizing antibody responses

Although Enterovirus 71 (EV71) has only one serotype based on serum neutralization tests using hyperimmune animal antisera, three major genogroups (A, B and C) including eleven genotypes (A, B1-B2, and C1-C5) can be well classified based on phylogenetic analysis. Since 1997, large-scale EV71 epidemics occurred cyclically with different genotypes in the Asia-Pacific region. Therefore, development of EV71 vaccines is a national priority in several Asian countries. Currently, five vaccine candidates have been evaluated in clinical trials in China (three C4 candidates), Singapore (one B2 candidate), and Taiwan (one B4 candidate). Overall, the peak viral titers of these 5 vaccine candidates could only reach about 10⁷ TCID₅₀/mL. Moreover, genotypes of these 5 candidates are different from the current predominant genotype B5 in Taiwan and South-Eastern Asia. We adapted a high-growth EV71 genotype B5 (HG-B5) virus after multiple passages and plaque selections in Vero cells and the HG-B5 virus could reach high titers (>10⁸ TCID₅₀/mL) in a microcarrier-based cell culture system. The viral particles were further purified and formulated with alum adjuvant. After two doses of intramuscular immunization in rabbits, the HG-B5 vaccine candidate could induce cross-reactive neutralizing antibodies against the three major EV71 genogroups. In conclusion, a high-growth EV71 virus was successfully adapted in Vero cells and could induce broad spectrum neutralizing antibody titers against three (A, B5, and C4) genotypes in rabbits.     

EV71 型脑炎患儿脑脊液和血清中 S-100b 蛋白 、VEGF 及I L-18 水平共同检测的临床意义

目的 探索脑脊液和血清中S-100b蛋白、VEGF、IL-18水平对肠道病毒71型(EV71)脑炎患儿的临床诊断意义.方法 60例EV71感染致重型脑炎患儿和40例轻型脑炎患儿为观察组 , 60例非中枢神经系统感染患儿为对照组.对急性期和恢复期EV71型脑炎患儿及正常对照组患儿脑脊液和血清中S-100b蛋白、VEGF及IL-18水平使用酶联免疫吸附试验( ELISA)法进行动态观察和对比分析.结果 重型EV71型脑炎患儿脑脊液和血清中S-100b蛋白、VEGF及IL-18水平显著高于轻型组EV71型脑炎患儿 ,差异有统计学意义( P <0.05).急性期EV71型脑炎患儿脑脊液和血清中S-100b蛋白、VEGF及IL-18水平显著高于恢复期,差异有统计学意义( P <0. 05).EV71型脑炎患儿脑脊液和血清中S-100b蛋白、VEGF及IL-18水平惊厥/抽搐组显著高于无惊厥/抽搐组,差异有统计学意义( P <0.05) ,意识障碍组显著高于无意识障碍组,差异有统计学意义( P <0.05).EV71 型脑炎患儿脑脊液和血清中S-100b蛋白、VEGF及IL-18水平表现出正相关性( r =0.012、0.168、0.968 ,P均<0.05).结论 脑脊液和血清中的S-100b蛋白、VEGF及IL-18水平可反映EV71感染损害脑组织的程度 ,能够评估预后 ,从而指导临床治疗.     

新型抗EV71病毒3C蛋白酶抑制药的设计、合成及其活性

摘 要 目的： 寻找抗EV71病毒新化合物。方法: 采用分子模拟技术,以EV71病毒3C蛋白酶活性口袋为靶,设计二肽、三肽,四肽,五肽和六肽等多个系列寡肽化合物,合成对接评分较高的寡肽化合物并进行抗EV71病毒活性筛选。结果: 五肽对接评分高于其他系列寡肽,显示五肽化合物与EV71 病毒3C蛋白酶有更强的键合力。研究中合成了两个评分较高的五肽化合物25和化合物16,体外抗EV71病毒活性实验显示化合物25活性较强（EC₅₀=1.36 μmol·L^-1,CC₅₀=211.94 μmol·L^-1, SI=155.84）,而化合物16则无明显活性。结论:五肽化合物25可作为抗EV71病毒新骨架开展进一步研究。     

EV71受体SCARB2和 PSGL-1在重症手足口病患者肺组织中的表达

目的：探讨人肠道病毒71型(enterovirus 71, EV71)清道夫受体B2(scavenger receptor class B member 2, SCARB2)和人P选择素糖蛋白配体-1(P-selectin glycoprotein ligand-1, PSGL-1)在重症手足口病(hand, foot and mouth disease, HFMD)患者、正常儿童及成人肺组织中的定位和分布。方法运用免疫组化SuperVision两步法检测15例重症HFMD患者、3例正常儿童及8例成人肺组织中SCARB2和PSGL-1的表达及分布。结果 SCARB2分布于重症HFMD患者、正常儿童及成人肺组织支气管上皮、细支气管上皮、肺泡上皮细胞及炎症细胞,其在三组中的阳性率差异无显著性( P>0．05)。 PSGL-1分布于成人肺组织支气管、细支气管上皮,但在重症HFMD患者及正常儿童中不表达,阳性率分别为100%、0、0,三组比较差异均有统计学意义(P<0．05),三组肺组织的炎症细胞PSGL-1均为阳性,阳性率分别为100%、66．7%、100%(P>0．05);PSGL-1在三组的肺泡上皮细胞中均不表达。结论 EV71受体SCARB2分布于重症HFMD患者肺组织支气管上皮、细支气管上皮、炎症细胞及肺泡上皮细胞,PSGL-1仅分布于重症HFMD患者肺组织炎症细胞,提示SCARB2可能在重症HFMD的感染过程中具有一定作用。     

Calcium deposition in photocrosslinked poly(Pro‐Hyp‐Gly) hydrogels encapsulated rat bone marrow stromal cells

Reproducing the features of the extracellular matrix is important for fabricating three‐dimensional (3D) scaffolds for tissue regeneration. A collagen‐like polypeptide, poly(Pro‐Hyp‐Gly), is a promising material for 3D scaffolds because of its excellent physical properties, biocompatibility, and biodegradability. In this paper, we present a novel photocrosslinked poly(Pro‐Hyp‐Gly) hydrogel as a 3D scaffold for simultaneous rat bone marrow stromal cell (rBMSC) encapsulation. The hydrogels were fabricated using visible‐light photocrosslinking at various concentrations of methacrylated poly(Pro‐Hyp‐Gly) (20–50 mg/ml) and irradiation times (3 or 5 min). The results show that the rBMSCs encapsulated in the hydrogels survived 7 days of incubation. Calcium deposition on the encapsulated rBMSCs was assessed with scanning electron microscope observation, Alizarin Red S, and von Kossa staining. The most strongly stained area was observed in the hydrogel formed with 30 mg/ml of methacrylated poly(Pro‐Hyp‐Gly) with 5‐min irradiation. These findings demonstrate that poly(Pro‐Hyp‐Gly) hydrogels support rBMSC viability and differentiation, as well as demonstrating the feasibility of using poly(Pro‐Hyp‐Gly) hydrogels as a cytocompatible, biodegradable 3D scaffold for tissue regeneration.     

Immunogenicity and performance of an enterovirus 71 virus-like-particle vaccine in nonhuman primates

A vaccine against human enterovirus 71 (EV-A71) is urgently needed to combat outbreaks of EV-A71 and in particular, the serious neurological complications that manifest during these outbreaks. In this study, an EV-A71 virus-like-particle (VLP) based on a B5 subgenogroup (EV-A71-B5 VLP) was generated using an insect cell/baculovirus platform. Biochemical analysis demonstrated that the purified VLP had a highly native procapsid structure and initial studies in vivo demonstrated that the VLPs were immunogenic in mice. The impact of VLP immunization on infection was examined in non-human primates using a VLP prime-boost strategy prior to EV-A71 challenge. Rhesus macaques were immunized on day 0 and day 21 with VLPs (100 μg/dose) containing adjuvant or with adjuvant alone (controls), and were challenged with EV-A71 on day 42. Complete blood counts, serum chemistry, magnetic resonance imaging (MRI) scans, and histopathology results were mostly normal in vaccinated and control animals after virus challenge demonstrating that the fatal EV-A71-B3 clinical isolate used in this study was not highly virulent in rhesus macaques. Viral genome and/or infectious virus were detected in blood, spleen or brain of two of three control animals, but not in any specimens from the vaccinated animals, indicating that VLP immunization prevented systemic spread of EV-A71 in rhesus macaques. High levels of IgM and IgG were detected in VLP-vaccinated animals and these responses were highly specific for EV-A71 particles and capsid proteins. Serum from vaccinated animals also exhibited similar neutralizing activity against different subgenogroups of EV-A71 demonstrating that the VLPs induced cross-neutralizing antibodies. In conclusion, our EV-A71-B5 VLP is safe, highly immunogenic, and prevents systemic EV-A71-B3 infection in nonhuman primates making it a viable attractive vaccine candidate for EV-A71.