壳聚糖-明胶/聚乳酸-羟基乙酸联合载药水凝胶的体外抗结核作用

文题释义：基质细胞衍生因子1：是一种参与免疫细胞活化、分化和迁移及伤口愈合、角膜上皮再生和组织修复等过程的趋化因子,能促进干细胞的生长和发育,参与调节成骨分化,可通过细胞归巢提高干细胞向病灶区的趋化作用。而基质细胞衍生因子1的失活会损害成骨细胞的发育和分化。此外,其还与血管生成密切相关。 异烟肼：具有较高的杀菌活性,是治疗结核病的一线药物。世卫组织建议将异烟肼作为结核病的标准疗法,用于潜伏性结核病感染者的预防治疗,与利福平、吡嗪酰胺和乙胺丁醇一起用于治疗活动性肺结核。异烟肼的活化形式与脂肪酸生物合成Ⅱ型系统中的NADH依赖型烯醇酰基载体蛋白还原酶异烟肼a结合,阻断细菌细胞壁关键成分支原体酸的合成。 背景：抗结核化疗是目前治疗骨关节结核的主要手段,然而全身给药难以维持病灶区的有效浓度,治疗效果欠佳。 目的：制备一种原位、长期释放抗结核药物且兼备促成骨作用的壳聚糖-明胶/聚乳酸-羟基乙酸联合载药水凝胶。 方法：将亲水性的抗结核药物异烟肼和疏水性的基质细胞衍生因子通过复乳法负载到聚乳酸-羟基乙酸中,制备聚乳酸-羟基乙酸载药微球,共混至壳聚糖-明胶水凝胶支架中,制备壳聚糖-明胶/聚乳酸-羟基乙酸联合载药水凝胶。检测聚乳酸-羟基乙酸载药微球、壳聚糖-明胶/聚乳酸-羟基乙酸联合载药水凝胶的体外释药与抗结核杆菌的能力。将成骨前体细胞MC3T3-E1分别接种于载药微球与联合载药水凝胶表面,CCK-8法检测细胞活力,碱性磷酸酶活性检测细胞的成骨性能。 结果与结论：①载药微球中异烟肼1 h内的突释约为23.3%,2 d内的释放率约为42.6%,随后进入缓释期,25 d后进入平台期;基质细胞衍生因子1在1 h内的累积释放率约为19.8%,2 d内的释放率约为44.7%,随后进入缓释期,25 d后进入平台期;联合载药水凝胶中异烟肼和基质细胞衍生因子1最初1 h的释放分别为8.3%和8.5%,第2天的累计释放率分别为15.2%和17.6%,远低于聚乳酸-羟基乙酸微球;②体外4周后,联合载药水凝胶的抑菌直径大于载药微球,抑菌率高于载药微球(P < 0.05);③联合载药水凝胶与载药微球均具有良好的细胞相容性,细胞活力均约为100%;④培养5,10 d后,联合载药水凝胶表面的细胞碱性磷酸酶活性与载药微球比较差异无显著性意义(P > 0.05);⑤结果表明,原位壳聚糖-明胶/聚乳酸-羟基乙酸联合载药水凝胶有作为治疗骨关节结核及其他骨关节感染的潜力。 ORCID: 0000-0003-4166-2492(张贺龙) 中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

肝靶向性壳聚糖基纳米载药体系的研究与应用

目的:阐述肝靶向性壳聚糖基纳米载药体系的研究和应用。方法:采用电子检索的方式,在万方数据库中检索2002-12/2010-02有关肝靶向性壳聚糖基纳米的研究文章,关键词为"壳聚糖基纳米载药体系,肝靶向性,研究,应用"。排除重复研究、普通综述或Meta分析类文章,筛选纳入18篇文献进行阐述。结果:壳聚糖纳米粒是一种天然无毒的非病毒药物载体,有良好的生物相容性和生物可降解性,可提高药物的稳定性,可靶向释放药物,增加药物的吸收等,达到控释和靶向治疗的作用。载药壳聚糖纳米粒的制备有离子交联法、沉淀法、超声乳化法、反相微乳法、静电纺丝法、反相悬浮交联法、逆相蒸发一短时超声法、还原胺化法等8种不同方法。用于肝脏组织工程如肝移植、人工肝,能维持和提高肝细胞活性和功能,有利于肝细胞生长;肝脏肿瘤治疗中使药物靶向释放于肿瘤部位,更有效抑制肿瘤细胞、降低毒副作用等。结论:肝靶向性壳聚糖基纳米载药体系是一种安全、高效的靶向性基因载体,但它的研究还需不断深入。     

温敏性壳聚糖水凝胶对大鼠成骨细胞的毒性

背景：温敏性壳聚糖与多种细胞相容性良好,是组织工程中不可多得的优良载体,但其对成骨细胞毒性研究相对缺乏。 目的：验证温敏性壳聚糖水凝胶对成骨细胞的毒性。 方法：成骨细胞在温敏性壳聚糖水凝胶中进行培养,显微镜下观察细胞形态及扩增情况,同时,SD大鼠成骨细胞在不同浓度的温敏性壳聚糖水凝胶浸提液中体外培养24,48,72,96 h,MTT法测定细胞相对增殖率,判断细胞毒性的级别。 结果与结论：SD大鼠成骨细胞在温敏性壳聚糖水凝胶中培养24 h内镜下观察呈圆形,48 h后开始伸出触角并扩增;温敏性壳聚糖水凝胶浸提液中培养的各组细胞在不同时间点相对增殖率在92%~112%之间,各浓度的温敏性壳聚糖水凝胶材料浸提液的细胞毒性均为0级或1级,完全符合生物材料的安全评价标准。     

壳聚糖水凝胶与星形胶质细胞体外生物相容性评价的实验研究

研究壳聚糖水凝胶材料与星形胶质细胞的体外生物相容性,初步探讨壳聚糖水凝胶作为神经组织工程支架材料的可行性.利用氯化壳聚糖、β-甘油磷酸钠和羟乙基纤雏素制备壳聚糖水凝胶,MTT法评价其细胞毒性;体外培养鉴定新生Wistar大鼠脑皮层星形胶质细胞;壳聚糖水凝胶与星形胶质细胞体外共培养,观察星形胶质细胞在材料上的生长;MTT...     

壳聚糖水凝胶与星形胶质细胞体外生物相容性评价的实验研究

研究壳聚糖水凝胶材料与星形胶质细胞的体外生物相容性,初步探讨壳聚糖水凝胶作为神经组织工程支架材料的可行性。利用氯化壳聚糖、β-甘油磷酸钠和羟乙基纤维素制备壳聚糖水凝胶,MTT法评价其细胞毒性;体外培养鉴定新生Wistar大鼠脑皮层星形胶质细胞;壳聚糖水凝胶与星形胶质细胞体外共培养,观察星形胶质细胞在材料上的生长;MTT法检测接种后1、35、、7d的细胞增殖度。体外成功制备壳聚糖水凝胶,该材料无细胞毒性;体外分离、培养得到状态良好的星形胶质细胞;体外星形胶质细胞在材料上培养呈星形,生长良好,有分支状突起形成;MTT结果表明,材料-细胞共培养组中的细胞增殖度明显高于单纯细胞组（P〈0.001）。壳聚糖水凝胶与星形胶质细胞在体外具有良好的生物相容性,有望作为神经组织工程支架材料。     

壳聚糖水凝胶微球模板的载肝素层层自组装微胶囊

背景：近年来,对抗凝血药物肝素的控释一直存在争议,静电层层自组装是微胶囊载药简单有效的新方法。 目的：制备壳聚糖水凝胶微球模板的载肝素层层自组装微胶囊,从而实现对抗凝血药物肝素的控释。 方法：采用硫酸钠沉淀的方法制备了具有正电荷的壳聚糖微球模板,通过层层自组装的方法装载抗凝血药物肝素。壳聚糖(CS)作为聚阳离子和肝素(Heparin, Hep)作为聚阴离子,在壳聚糖微球的模板上层层自组装形成{CS/Hep}3。{CS/Hep}3包被壳聚糖微球模板的微胶囊通过倒置荧光显微镜、激光共聚焦显微镜和激光粒度分析进行了表征。壳聚糖和肝素的层层自组装过程通过Zeta电位分析进行监测。 结果与结论：{CS/Hep}3包被壳聚糖微球模板的微胶囊平均直径1 μm,包封率和载肝素量分别为83.8%和3.05%。成功制备了壳聚糖水凝胶微球模板的载肝素层层自组装微胶囊,进而实现对抗凝血药物肝素的控释。     

温敏型可注射水凝胶的制备研究

为得到应用于软组织重建的新型可注射组织工程支架材料,将经交联合成的HA交联凝胶(XLHA),分别与降解性能不同的两种温敏型材料聚N-异丙基丙烯酰胺(PNIPAAm)凝胶和甲基纤维素(MC),在常温下共混合成XLHA-PNIPAAm与XLHA-MC可注射水凝胶,分析XLHA-PNIPAAm与XLHA-MC水凝胶的可注射性能、耐酶解性能、温敏性能和化学结构,并进行溶血试验与细胞毒性试验评价可注射凝胶的生物相容性。研究获得了XLHA-PNIPAAm的可注射水凝胶的制备条件,并证实了分别将不可降解材料PNIPAAm与可降解材料MC和HA共混均能有效延缓HA的降解,且PNIPAAm延缓HA降解的效果更为显著。辐照剂量为5 kGy,MBAAm/NIPAAm(M/M)=0.015,NIPAAm单体浓度为3%,制备的PNIPAAm与XLHA共混制备的XLHA-PNIPAAm可注射凝胶的耐酶解性能最佳。差示扫描量热仪(DSC)检测显示XLHA-PNIPAAm的温敏性较为稳定。两种复合凝胶的细胞毒性和溶血试验均合格,且XLHA-PNIPAAm水凝胶的生物相容性要好于XLHA-MC凝胶。     

银离子水凝胶涂层导尿管抑菌效力定量检查方法的建立

目的建立银离子水凝胶涂层导尿管的抑菌效力定量检查方法。方法根据《中华人民共和国药典》2015年版微生物计数法(四部通则1105),以金黄色葡萄球菌、铜绿假单胞菌、大肠埃希菌、表皮葡萄球菌和白色念珠菌作为试验菌株,以D/E中和肉汤作为中和剂,采用中和剂与薄膜过滤法联合使用的方式对银离子水凝胶涂层导尿管进行微生物计数方法适用性试验。根据《中华人民共和国药典》2015年版抑菌效力检查法(四部通则1121),采用经验证的微生物计数方法定量测定银离子水凝胶涂层导尿管对上述5种试验菌的抑菌效力。结果D/E中和肉汤与薄膜过滤法联合使用可以有效中和银离子的抑菌活性,银离子水凝胶涂层导尿管对上述5种试验菌均具有较好的抑菌作用,抑菌率均能达到99%,且抑菌效力可持续至14 d。结论采用D/E中和肉汤与薄膜过滤法联合使用的方式建立适用于银离子水凝胶涂层导尿管的微生物计数方法,可照此方法进行抑菌效力检查中存活菌数的测定。建立的方法可用于银离子水凝胶涂层导尿管抑菌效力的定量检查。     

纳米羟基磷灰石/阿司匹林/聚乙烯醇/明胶/海藻酸钠水凝胶支架的制备与表征

背景：阿司匹林是经典非类固醇抗炎药物,合适剂量的阿司匹林可调节免疫、促进成骨。制备出可缓释阿司匹林、调控骨缺损区免疫微环境和加速骨缺损修复的骨组织工程材料是目前的研究热点。目的：制备可调控免疫微环境和加速骨缺损修复的新型水凝胶支架。方法：制备含有质量分数0%,10%和20%纳米羟基磷灰石的纳米羟基磷灰石/阿司匹林/聚乙烯醇/明胶/海藻酸钠水凝胶支架,对水凝胶支架的微观结构、孔隙率、化学成分、晶相结构、药物缓释性能、力学性能、溶胀性能和降解性能进行表征。通过细胞增殖和细胞毒性实验评价水凝胶支架的生物相容性。结果与结论：(1)扫描电镜和孔隙率检测结果表明,0%,10%纳米羟基磷灰石组水凝胶支架具有更仿生的层级多孔结构、孔隙通联性和孔隙率;(2)傅里叶红外光谱和X射线衍射结果表明,水凝胶支架内各原料以物理、化学双交联方式结合,且不会破坏纳米羟基磷灰石的晶相结构;(3)力学性能结果表明,10%纳米羟基磷灰石组水凝胶支架具有最佳的压缩模量和压缩强度;(4)药物释放性能结果显示,随着纳米羟基磷灰石质量的增加,水凝胶支架的阿司匹林累计释放率降低,但是药物突释率降低、缓释时间延长;(5)溶胀性能和降解...     

骨髓间充质干细胞复合温敏水凝胶支架的制备

背景：壳聚糖及其衍生物制备的支架对细胞迁移和神经轴突再生有重要作用。壳聚糖及其衍生物的组织相容性好,易使干细胞在其表面附着生长,在神经组织工程具有较为广阔的应用前景。 目的：制备适宜骨髓间充质干细胞生长的壳聚糖/壳聚糖季铵盐/甘油磷酸钠温敏性水凝胶细胞支架,观察骨髓间充质干细胞在细胞支架中的生长情况。 方法：将壳聚糖进行季铵盐化改性处理,通过傅里叶变换红外光谱分析谱检测确定其生成。实验以壳聚糖与壳聚糖季铵盐配比为8∶1成功制备出较为稳定的壳聚糖/壳聚糖季铵盐/甘油磷酸钠温敏性温敏水凝胶细胞支架,观察成胶情况,并进行生物安全性检测。 结果与结论：实验在傅里叶变换红外光图谱上发现了季铵基基团的特征峰。细胞毒性实验显示,水凝胶浸提液干预的大鼠骨髓间充质干细胞无毒性。急性全身毒性实验显示,浸提液对大鼠体质量增加无明显影响,支架生物安全性较好。扫描电镜观察显示,骨髓间充质干细胞在细胞支架中能正常的生长和增殖。结果证实,实验成功制备了壳聚糖/壳聚糖季铵盐/甘油磷酸钠温敏性水凝胶细胞支架,适合骨髓间充质干细胞生长和增殖。     

Renal dysplasia and asplenia in two sibs

A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.     

Schizophrenia in a North Swedish geographical isolate, 1900–1977. Epidemiology, genetics and biochemistry

Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc² (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.     

The dominant diseases of modernized societies as omega-3 essential fatty acid deficiency syndrome: Substrate beriberi

About 1900, modern food selection and processing caused widespread $\text{epidemics}$ of the B vitamin deficiency diseases of beriberi and pellagra which, for genetic reasons, often expressed as different diseases ranging from bowel and heart disease to dermatoses and psychoses. But the B vitamins merely help convert essential fatty acids (EFA) into the prostaglandin (PG) tissue regulators and it now turns out that, through hydrogenation, milling and selection of w3-poor southern foods, we have also been systematically depleting, by as much as 90%, a newly discovered trace Nordic EFA (w3) of special importance to primates and sole precursor of the PG3(4) series, even as a concurrent fiber deficiency increases body demand for EFA. Since substrate EFA is processed by many B vitamin catalysts, an EFA deficiency will mimic a $\text{panh}$ypovitaminosis B, i.e., a mixture of $\text{substrate}$ beriberi and $\text{substrate}$ pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse $\text{endemic}$ disease. This would consitute a second stage of the Modern Malnutrition and explain why some workers now hold the dominant diseases of modermized societies to be new, nutritionally based, pellagraform yet lipid-related and to range, once again, from heart disease to psychosis. It is an assumption that our dominant diseases are unrelated to each other or are merely revealed by our diagnostic acumen and therapeutic success; and that hydrogenating millions of tons of food oils annually, to destroy the rancidity producing w3-EFA, is safe for $\text{primates}$. Extensive beriberiform disease is reported here in 32 typical cases taken from medical practice which responds strikingly to linseed oil supplements (60% w3-EFA) in confirmation of identical results in Capuchins.     

What will studies of Fulani individuals naturally exposed to malaria teach us about protective immunity to malaria?

There are an estimated over 200 million yearly cases of malaria worldwide. Despite concerted international effort to combat the disease, it still causes approximately half a million deaths every year, the majority of which are young children with Plasmodium falciparum infection in sub-Saharan Africa. Successes are largely attributed to malaria prevention strategies, such as insecticide-treated mosquito nets and indoor spraying, as well as improved access to existing treatments. One important hurdle to new approaches for the treatment and prevention of malaria is our limited understanding of the biology of Plasmodium infection and its complex interaction with the immune system of its human host. Therefore, the elimination of malaria in Africa not only relies on existing tools to reduce malaria burden, but also requires fundamental research to develop innovative approaches. Here, we summarize our discoveries from investigations of ethnic groups of West Africa who have different susceptibility to malaria.     

'Very sore nights and days': the child's experience of illness in early modern England, c.1580-1720

Sick children were ubiquitous in early modern England, and yet they have received very little attention from historians. Taking the elusive perspective of the child, this article explores the physical, emotional, and spiritual experience of illness in England between approximately 1580 and 1720. What was it like being ill and suffering pain? How did the young respond emotionally to the anticipation of death? It is argued that children’s experiences were characterised by profound ambivalence: illness could be terrifying and distressing, but also a source of emotional and spiritual fulfilment and joy. This interpretation challenges the common assumption amongst medical historians that the experiences of early modern patients were utterly miserable. It also sheds light on children’s emotional feelings for their parents, a subject often overlooked in the historiography of childhood. The primary sources used in this article include diaries, autobiographies, letters, the biographies of pious children, printed possession cases, doctors’ casebooks, and theological treatises concerning the afterlife.     

Guidelines for the Validation and Use of Immunoassays for Determination of Introduced Proteins in Biotechnology Enhanced Crops and Derived Food Ingredients

Recent advancements in agricultural biotechnology have created a need for analytical techniques to determine introduced proteins in crops enhanced through modern biotechnology techniques. These proteins are expressed in plant tissues and may be present in food ingredients. Immunoassays are ideally suited for protein detection and may be used as both quantitative and threshold methods. Microplate ELISA and lateral flow devices are two of the most commonly used immunoassay formats for agricultural biotechnology applications. This paper provides general background information and a discussion of criteria for the validation and application of immunochemical methods to the analysis of proteins introduced into plants and food ingredients using biotechnology methods. It is the result of a collaborative effort of members of the Analytical Environmental Immunochemical Consortium. This collaborative effort represents the combined expertise of several organizations to reach consensus on establishing guidelines for the validation and use of immunoassays. Further, the paper offers developers and users a consistent approach to adopting the technology as well as aid in producing accurate and meaningful results.     

Ultracryotomy: development and application (with examples from the yeast cytology) (author's transl)

The preparation steps usually necessary for obtaining ultrathin frozen sections of biological material (chemical prefixation, enclosing, cryoprotective treatment, freezing, sectioning, and post-staining the sections for transmission electron microscopy) are submitted to a critical analysis. The application of cryo-ultramicrotomy, in particularly for cytochemical purposes, is reviewed. Fundamental considerations of chemical prefixation and poststaining are supported by examples from yeast cytology. Furthermore, the efficiency of the cryo-ultramicrotomy (electron optical resolution of ultrastructural details) is demonstrated on yeast cells and protoplasts.     

HLA in North Colombia Chimila Amerindians

HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.     

The universal muscular chamber. A basic unit of the genitourinary, cardiovascular, gastro-intestinal, skeletal, cutaneous, respiratory and other systems, endocameral hypertension; and spasm, the key to their idiopathic diseases and arthropathies

Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.