Use of ex vivo and in vitro cultures of the human respiratory tract to study the tropism and host responses of highly pathogenic avian influenza A (H5N1) and other influenza viruses

The tropism of influenza viruses for the human respiratory tract is a key determinant of host-range, and consequently, of pathogenesis and transmission. Insights can be obtained from clinical and autopsy studies of human disease and relevant animal models. Ex vivo cultures of the human respiratory tract and in vitro cultures of primary human cells can provide complementary information provided they are physiologically comparable in relevant characteristics to human tissues in vivo, e.g. virus receptor distribution, state of differentiation. We review different experimental models for their physiological relevance and summarize available data using these cultures in relation to highly pathogenic avian influenza H5N1, in comparison where relevant, with other influenza viruses. Transformed continuous cell-lines often differ in important ways to the corresponding tissues in vivo.     

Cyclic mechanical stimulation rescues achilles tendon from degeneration in a bioreactor system

Physiotherapy is one of the effective treatments for tendinopathy, whereby symptoms are relieved by changing the biomechanical environment of the pathological tendon. However, the underlying mechanism remains unclear. In this study, we first established a model of progressive tendinopathy‐like degeneration in the rabbit Achilles. Following ex vivo loading deprivation culture in a bioreactor system for 6 and 12 days, tendons exhibited progressive degenerative changes, abnormal collagen type III production, increased cell apoptosis, and weakened mechanical properties. When intervention was applied at day 7 for another 6 days by using cyclic tensile mechanical stimulation (6% strain, 0.25 Hz, 8 h/day) in a bioreactor, the pathological changes and mechanical properties were almost restored to levels seen in healthy tendon. Our results indicated that a proper biomechanical environment was able to rescue early‐stage pathological changes by increased collagen type I production, decreased collagen degradation and cell apoptosis. The ex vivo model developed in this study allows systematic study on the effect of mechanical stimulation on tendon biology. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 33:1888–1896, 2015.     

Stem cell bioengineering strategies to widen the therapeutic applications of haematopoietic stem/progenitor cells from umbilical cord blood

Umbilical cord blood (UCB) transplantation has observed a significant increase in recent years, due to the unique features of UCB haematopoietic stem/progenitor cells (HSCs) for the treatment of blood‐related disorders. However, the low cell numbers available per UCB unit significantly impairs the widespread use of this source for transplantation of adult patients, resulting in graft failure, delayed engraftment and delayed immune reconstitution. In order to overcome this issue, distinct approaches are now being considered in clinical trials, such as double‐UCB transplantation, intrabone injection or ex vivo expansion. In this article the authors review the current state of the art, future trends and challenges on the ex vivo expansion of UCB HSCs, focusing on culture parameters affecting the yield and quality of the expanded HSC grafts: novel HSC selection schemes prior to cell culture, cytokine/growth factor cocktails, the impact of biochemical factors (e.g. O₂) or the addition of supportive cells, e.g. mesenchymal stem/stromal cell (MSC)‐based feeder layers) were addressed. Importantly, a critical challenge in cellular therapy is still the scalability, reproducibility and control of the expansion process, in order to meet the clinical requirements for therapeutic applications. Efficient design of bioreactor systems and operation modes are now the focus of many bioengineers, integrating the increasing 'know‐how' on HSC biology and physiology, while complying with the GMP standards for the production of cellular products, i.e. through the use of commercially available, highly controlled, disposable technologies. Copyright © 2013 John Wiley & Sons, Ltd.     

覆盆子活性成分山奈酚的含量测定

目的：建立覆盆子改善肾阳虚型痴呆活性成分山奈酚的含量测定方法。方法：采用高效液相色谱法。色谱柱为Diamon－sil C18（250mm ×4.6mm,5μm）,流动相：乙腈－0.1％磷酸水（30∶70）,流速1 mL／min,柱温30℃,检测波长266nm,进样量20μL。结果：山奈酚质量在0.0782～5.000μg范围内与峰面积呈良好的线性关系（r＝0.9999）,平均加样回收率为99.50％, RSD＝2.1％。结论：试验方法简便、快速、实验结果准确可靠,该方法可作为覆盆子有效成分山奈酚含量测定的方法。     

光照对粗茎秦艽种子萌发及其过程中环烯醚萜类成分积累规律的影响

目的:观察光照对粗茎秦艽种子萌发及其过程中环烯醚萜类成分积累规律的影响。方法:将粗茎秦艽种子分为光照组和避光组,浸泡24 h后置于21℃培养箱内培养,光照组的光照强度为605～710 lx。以种皮开裂露出胚根为发芽标志,记录每日种子的发芽情况。培养33 d后,分别统计光照组和避光组的种子发芽率、首次萌发时间与半数萌发时间,测量其子叶、胚轴、胚根的长度及胚轴直径。于培养过程中不同时间点取样,液质联用技术定性鉴别环烯醚萜类化合物,并对各成分含量进行相对定量分析。比较光照组和避光组种子萌发过程中环烯醚萜类成分含量的变化趋势。结果:①培养33 d后,光照组平均发芽率为76%,避光组平均发芽率为88%;光照组种子首次萌发时间略晚于避光组;光照组和避光组的半数萌发时间分别为培养16、15 d。②避光组子叶未见发育,光照组子叶发育明显。避光组的胚轴长度和胚轴直径明显大于光照组(P0.01),光照组的胚根长度明显大于避光组(P0.01)。③液质联用定性鉴别出8种环烯醚萜类化合物。种子萌发过程中,龙胆苦苷含量逐渐增加,而马钱苷酸含量逐渐减少;环烯醚萜衍生物gentistraminosides B、macrophylloside A、macrophylloside B及其同分异构体含量逐渐增加。光照组的龙胆苦苷和以上4种环烯醚萜衍生物的含量高于避光组。结论:光照对粗茎秦艽种子萌发及其胚轴伸长具有一定抑制作用,并对萌发过程中环烯醚萜类成分的积累产生影响。     

A Case of Carcinoma Ex Pleomorphic Adenoma in the Buccal Mucosa: Review of the Literature

We present a case of carcinoma ex pleomorphic adenoma on the right buccal mucosa in a 52-year-old Japanese woman. Based on the histopathology, the excised tumor was the non-invasive type, but the majority of the tumor consisted of poorly-differentiated adenocarcinoma cells. We performed proton radiation after the surgery. The patient was well, without evidence of disease, 48 months after surgery. Carcinoma ex pleomorphic adenoma in the buccal mucosa has been reported in only four cases during the past twenty years. Therefore, our case was comparatively rare.     

改良运动处方对慢性阻塞性肺疾病稳定期患者的康复疗效

目的 以联合缩唇-腹式呼吸法和六字诀训练为改良的运动处方,探讨其对慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)稳定期患者的康复疗效.方法 将100例患者随机分为四组:对照组、缩唇-腹式呼吸组、六字诀组、运动处方组,每组25例,各组分别于锻炼前和坚持锻炼半年后测定肺功能、6分钟步行距离(6MWD)和动脉血气分析.结果 运动处方组较对照组、缩唇-腹式呼吸组、六字诀组的肺功能指标FEV1、FEV1占预计值百分比和6MWD及血气分析等有明显改善,差异有统计学意义.结论 改良运动处方能有效改善COPD稳定期患者的肺功能、提高运动耐力、提高动脉血氧分压(PaO2)及降低动脉二氧化碳分压(PaCO2),且在改善肺功能、提高运动耐力及降低PaCO2等方面的疗效明显优于单独的缩唇-腹式呼吸操和六字诀训练.     

Simple Analysis of the Computed Tomography Features of Gastric Schwannoma

ObjectiveThe objective of this study was to assess the computed tomography (CT) findings of gastric schwannoma (GS) and identify the difference between large (> 5 cm) and small (≤ 5 cm) GS.Materials and MethodsCT findings of 38 pathologically proven cases of GSs were retrospectively reviewed. The CT evaluation of GS included categorical variables (location, contour, growth pattern, enhancement pattern, necrosis, ulceration, calcification, and lymph nodes) and continuous variables (size, CT value of 3 phases, and enhancement degree). The lesion was divided into 2 groups (large [> 5 cm] and small [≤ 5 cm] GS) according to the tumor size. The Fisher exact test was used for categorical variables and the Student t or Mann-Whitney U test for continuous variables.ResultsOf the 38 patients, there were 32 women and 6 men. The median age was 54.5 years (range 39–79). Most of patients (65.8%, [25 of 38]) had nonspecific gastrointestinal symptoms such as abdominal or gastric pain, fullness and discomfort, bleeding, and melena. The tumors were mainly located in the stomach body (71.1% [27 of 38]), and the mean diameter was 3.7 cm (range 1.5 cm-10.3 cm), of which included large (> 5 cm) (n = 8) and small (≤ 5 cm) (n = 30). All of the GSs were benign, 9 of whom had palpable perigastric lymph nodes, which confirmed by pathology for the reactive inflammatory hyperplasia. Growth pattern, pattern of enhancement, necrosis, calcification, surface ulceration, and lymph node in the CT images were found to be significant variables for differentiating large (> 5 cm) and small (≤ 5 cm) GS (P < .05).ConclusionGSs were predominantly located at the gastric body and occurred most frequently in women between the ages of 40–70 years, and showed gradual enhancement after contrast enhancement. Palpable perigastric lymph nodes could not be considered as malignant factor of GS. There 7 computed CT criteria are significant difference between large (> 5 cm) and small (≤ 5 cm) GS.