64例儿童门静脉高压肝脏病理学分析

目的探讨儿童门静脉高压的临床病理特征。方法对64例儿童门静脉高压的临床情况及肝组织病理形态进行分析。结果64例儿童门静脉高压患儿年龄从1岁3个月至14岁,平均年龄6.3岁。病程2 d至8年,临床主要表现为消化道出血50例(78.1%);脾肿大59例;肝肿大5例;发热8例。B超及CT检查提示门静脉海绵变性57例(89.1%);脾大59例(92.2%);8例提示肝硬化。病理学检查,57例门静脉海绵变性肝脏组织主要表现为门静脉高压的压力相关性形态学改变,其中6例合并其他肝脏病变。7例肝性门静脉高压中包含先天性肝纤维化2例,小结节性肝硬化2例,肝外胆道闭锁致胆汁性肝硬化1例,结节再生性增生1例,非硬化性门静脉高压1例。54例获得随访。结论儿童门静脉高压病因与成人不同,肝组织活检是确诊的重要手段。     

High-coverage SARS-CoV-2 genome sequences acquired by target capture sequencing

In this study, we designed a set of SARS-CoV-2 enrichment probes to increase the capacity for sequence-based virus detection and obtain the comprehensive genome sequence at the same time. This universal SARS-CoV-2 enrichment probe set contains 502 120 nt single-stranded DNA biotin-labeled probes designed based on all available SARS-CoV-2 viral sequences and it can be used to enrich for SARS-CoV-2 sequences without prior knowledge of type or subtype. Following the CDC health and safety guidelines, marked enrichment was demonstrated in a virus strain sample from cell culture, three nasopharyngeal swab samples (cycle threshold [C_t] values: 32.36, 36.72, and 38.44) from patients diagnosed with COVID-19 (positive control) and four throat swab samples from patients without COVID-19 (negative controls), respectively. Moreover, based on these high-quality sequences, we discuss the heterozygosity and viral expression during coronavirus replication and its phylogenetic relationship with other selected high-quality samples from the Genome Variation Map. Therefore, this universal SARS-CoV-2 enrichment probe system can capture and enrich SARS-CoV-2 viral sequences selectively and effectively in different samples, especially clinical swab samples with a relatively low concentration of viral particles.     

新型3D打印导板辅助颈椎前路双侧椎弓根置钉的安全及可行性

文题释义：3D 打印手术导板：该导板依据手术需要,通过计算机辅助设计、3D打印制备出的一种具有术中准确定位点、置钉的位置、方向和深度,精确建立孔道、截面、空间距离、相互成角关系及其他复杂空间结构等功能的辅助手术器械,具有获取途径方便、使用方法简单、价格低廉等优点。 颈椎前路椎弓根置钉：是一种区别于传统颈椎后路椎弓根螺钉置钉技术的新型置钉技术,该技术可通过单独前方入路实现颈椎三柱损伤或多节段病变的坚强固定,无需再行后路固定,可有效减少手术创伤和并发症。 背景：下颈椎前路椎弓根螺钉固定技术可通过前方入路实现颈椎坚强固定,具有较大的应用前景。但该技术操作难度大、风险高,目前尚未得到广泛应用。 目的：改良设计一种用于下颈椎双侧前路椎弓根螺钉置钉的3D打印导板,探讨其辅助颈椎前路椎弓根螺钉置钉的可行性及安全性。 方法：选取6具正常成人颈椎标本,男女各3具,行薄层CT扫描后将影像数据以DICOM格式导入Mimics 17.0软件,三维重建后模拟设计出C₃-C₇双侧颈椎前路椎弓根螺钉钉道导孔,再以颈椎椎体前面、椎体上面前1/2及双侧钩突关节面前1/2骨性结构为标志,反向增厚设计生成导孔基座,形成颈椎前路椎弓根螺钉置钉导板。经3D打印得到导板实体后,在导板辅助下行C₃-C₇双侧颈椎前路椎弓根螺钉置钉。将置钉后的颈椎标本再次行CT扫描,通过CT断面影像评价置钉准确性。同时利用Mimics17.0软件比较实际钉道与模拟钉道在横断面的内、外偏移角度(α1、α2)差异及其在矢状面的上、下偏移角度(β1、β2)差异。 结果与结论：①双侧共计60枚颈椎前路椎弓根螺钉均顺利置入,其中57枚完全位于椎弓根皮质内,判定为0级,准确率95.0%;另外3枚破出椎弓根皮质,其中1级2枚(3.3%),2级1枚(1.7%);②真实钉道与模拟钉道相比,其横断面内、外偏移角分别为(0.867±0.787)°、(0.783±0.792)°,差异无显著性意义(P > 0.05);矢状面上、下偏移角分别为(1.362±1.380)°、(1.314±1.300)°,差异无显著性意义(P > 0.05)。③提示在设计得到的3D打印导板辅助下可顺利完成下颈椎双侧前路椎弓根螺钉置钉,且具有良好的置钉安全性。 ORCID: 0000-0003-0124-7585(肖强)中国组织工程研究杂志出版内容重点：人工关节;骨植入物;脊柱;骨折;内固定;数字化骨科;组织工程     

组织玻璃化冻存技术：优势与尚未完全解决的问题

文题释义：玻璃化冷冻保存：是利用这些高浓度的低温保护剂组合成玻璃化冻存液,通过与水分子发生强烈的水合作用,增加溶液黏性,降低冰晶形成速度,从而使细胞在快速降温或复温过程中得以保护。 玻璃化：对于非晶高分子,当高分子通过降温从高弹态转变为玻璃态,或者通过升温从玻璃态转变为高弹态的过程称之为玻璃化转变,发生玻璃化转变的温度叫玻璃化转变温度。对于结晶高分子,玻璃化转变是指其非晶部分所发生的由高弹态向玻璃态(或者玻璃态向高弹态)的转变。因此,玻璃化转变是高分子中普遍存在的现象。但是玻璃化转变现象并不局限于高分子,一些小分子化合物也存在玻璃化转变。 背景：玻璃化冷冻保存是一种应用前途广阔的低温冷冻方法,通过使用高浓度的玻璃化冻存试剂将生物材料进行玻璃态转变,从而实现活性保存。 目的：就玻璃化冻存的生物学原理及玻璃化冻存试剂的分类,卵巢、皮肤与角膜等医学组织标本的玻璃化冻存进行综述。 方法：以“tissue;vitrification;cryopreservation”为英文检索词;“组织;玻璃化;冷冻保存”为中文检索词,检索1994年1月至2019年10月 PubMed 数据库及万方医学网相关文献。按照纳入与排除标准筛选后,对最终纳入的45篇文献进行归纳总结。 结果与结论：玻璃化冻存可以防止细胞内外冰晶形成,避免了冰晶给细胞带来的多种损伤,有效保留了细胞的生物活性与基本功能。玻璃化冻存试剂主要分为渗透性和非渗透性2种,其操作简便、高效,唯一的缺点是高浓度的冻存试剂对细胞具有一定的毒性损伤。为了降低对组织整体损伤风险,可以混合使用多种低毒冻存试剂。目前玻璃化冻存技术已经成功应用于多种细胞,但组织冻存的技术难题尚未完全解决。 ORCID: 0000-0003-0140-9935(张源) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

牛膝甾酮干预SD乳鼠成骨细胞的增殖与分化

文题释义：牛膝甾酮：牛膝为苋科植物牛膝的干燥根,主要含有甾酮类、皂苷类及多糖类成分,具有调节糖代谢、抗炎、镇痛、降血糖、免疫调节等作用,牛膝甾酮是其活性成分之一,目前尚无文献报道其对成骨细胞的生物学效应。 成骨细胞：由间充质干细胞分化而来,直接参与骨形成,其增殖、分化缺陷是骨质疏松症发病的重要原因,研究表明改善成骨细胞功能可有效治疗骨质疏松症。 背景：促进成骨细胞的增殖以及分化是治疗骨质疏松症的有效手段之一,但关于牛膝甾酮对成骨细胞增殖、分化的影响却没有报道。 目的：研究牛膝甾酮对SD乳鼠成骨细胞增殖、分化的影响及其潜在的分子机制。 方法：通过酶消化法获得SD乳鼠成骨细胞,体外诱导培养并鉴定;采用CCK8法检测不同质量浓度牛膝甾酮(1,5,10 mg/L)对成骨细胞活力的影响;采用碱性磷酸酶染色及碱性磷酸酶活性试剂盒评价成骨细胞的早期分化能力;采用茜素红染色观察钙结节的形成数量,评估成骨细胞的矿化能力;采用实时荧光定量PCR法检测成骨分化标志物的表达水平;采用MDC染色检测自噬小体数量。 结果与结论：①相对于不加药对照组,牛膝甾酮在一定程度上抑制成骨细胞的增殖(P < 0.05),但却能够显著促进碱性磷酸酶活性及矿化结节的形成(P < 0.05),同时上调成骨分化标志物CollagenⅠ、OPG、OPN、OCN的表达水平,此外牛膝甾酮还能够促进自噬小体的形成;②结果提示,牛膝甾酮能够通过上调成骨分化相关基因及刺激自噬体的形成而促进成骨细胞的分化。 ORCID: 0000-0002-2837-6347(姜涛) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

Evaluation of Ion Torrent next-generation sequencing for thalassemia diagnosis

IntroductionTo evaluate a next-generation sequencing (NGS) workflow in the screening and diagnosis of thalassemia.MethodsIn this prospective study, blood samples were obtained from people undergoing genetic screening for thalassemia at our centre in Guangzhou, China. Genomic DNA was polymerase chain reaction (PCR)-amplified and sequenced using the Ion Torrent system and results compared with traditional genetic analyses.ResultsOf the 359 subjects, 148 (41%) were confirmed to have thalassemia. Variant detection identified 35 different types including the most common. Identification of the mutational sites by NGS were consistent with those identified by Sanger sequencing and Gap-PCR. The sensitivity and specificities of the Ion Torrent NGS were 100%. In a separate test of 16 samples, results were consistent when repeated ten times.ConclusionOur NGS workflow based on the Ion Torrent sequencer was successful in the detection of large deletions and non-deletional defects in thalassemia with high accuracy and repeatability.     

Haemorrhagic cystitis following the administration of voriconazole in the treatment of central nervous system aspergillosis: a case report

Central nervous system aspergillosis (CNS-A) is a rare and fatal fungal infection. Voriconazole is the recommended treatment for CNS-A. The therapeutic effect of voriconazole is good, but its use is limited due to adverse reactions. This case report describes a 37-year-old male patient that had previously been diagnosed with acute lymphoblastic leukaemia. He had received immunosuppressive agents for 1 year following a haematopoietic bone marrow transplant. He presented with a 1-month history of left limb weakness as well as recurrent fever. Brain magnetic resonance imaging showed that he had multiple cerebral infarctions. Subsequently, he was diagnosed with CNS-A by metagenomic next-generation sequencing. Voriconazole was added to his treatment regimen, but it resulted in severe haemorrhagic cystitis and possibly bladder rupture. The dose of voriconazole was adjusted and reparative bladder surgery was undertaken immediately. Eventually, the patient was successfully treated with voriconazole and there was no recurrence of symptoms after 1 year of follow-up. Haemorrhagic cystitis is a rare adverse drug reaction associated with voriconazole use. Based on the experience with this current case, physicians should be aware of urinary tract complications with voriconazole including haemorrhagic cystitis.     

Nanoparticle-directed sub-cellular localization of doxorubicin and the sensitization breast cancer cells by circumventing GST-Mediated drug resistance

Resistance to single or multiple chemotherapeutic drugs is a major complication in clinical oncology and is one of the most common treatment limitations in patients with reoccurring cancers. Nanoparticle (NP)-based drug delivery systems (DDS's) have been shown to overcome drug resistance in cancer cells mainly by avoiding the activation of efflux pumps in these cells. We demonstrate in this work that polyester-based hyperbranched dendritic-linear (HBDL)-based NPs carrying doxorubicin (Dox) can effectively overcome microsomal glutathione transferase 1 (MGST1)-mediated drug resistance in breast cancer cells. Our DDS was much more effective at considerably lower intracellular Dox concentrations (IC₅₀ 6.3 μm vs. 36.3 μm) and achieved significantly greater reductions in viability and induced higher degrees of apoptosis (31% vs. 14%) compared to the free drug in the resistant cells. Dox-loaded HBDL NPs were found to translocate across the membranes of resistant cells via active endocytic pathways and to be transported to lysosomes, mitochondria, and the endoplasmic reticulum. A significantly lower amount of Dox accumulated in these cytoplasmic compartments in resistant cells treated with free Dox. Moreover, we found that Dox-HBDL significantly decreased the expression of MGST1 and enhanced mitochondria-mediated apoptotic cell death compared to free Dox. Dox-HBDL also markedly activated the JNK pathway that contributes to the apoptosis of drug-resistant cells. These results suggest that HBDL NPs can modulate subcellular drug distribution by specific endocytic and trafficking pathways and that this results in drug delivery that alters enzyme levels and cellular signaling pathways and, most importantly, increases the induction of apoptosis. Our findings suggest that by exploiting the cell transport machinery we can optimize the polymeric vehicles for controlled drug release to overcome drug resistance combat drug resistance with much higher efficacy.