油茶花苞采集机械手的设计与试验

丘陵地区油茶林地众多,油茶花苞采集暂无机械化设备,故设计了一款六自由度的油茶花苞采集机械手。通过油茶花苞生物学特性分析,使用旋扭式末端执行器可以实现油茶花苞的高效采集,采净率为90%。通过机械手工作空间分析,确定了机械手的尺寸,并将末端执行器搭载到机械手上进行了油茶林田间试验。试验结果表明:搭载了旋扭式末端执行器六自由度机械手,可以到达采摘区间的边界位置,末端在x、y、z方向的最大偏差分别为3.4、3.3、3.4 cm,为油茶花苞智能化采集提供了依据。

A new horizon for neuroscience:terahertz biotechnology in brain research

Terahertz biotechnology has been increasingly applied in various biomedical fields and has especially shown great potential for application in brain sciences.In this article,we review the development of terahertz biotechnology and its applications in the field of neuropsychiatry.Available evidence indicates promising prospects for the use of terahertz spectroscopy and terahertz imaging techniques in the diagnosis of amyloid disease,cerebrovascular disease,glioma,psychiatric disease,traumatic brain injury,and myelin deficit.In vitro and animal experiments have also demonstrated the potential therapeutic value of terahertz technology in some neuropsychiatric diseases.Although the precise underlying mechanism of the interactions between terahertz electromagnetic waves and the biosystem is not yet fully understood,the research progress in this field shows great potential for biomedical noninvasive diagnostic and therapeutic applications.However,the biosafety of terahertz radiation requires further exploration regarding its two-sided efficacy in practical applications.This review demonstrates that terahertz biotechnology has the potential to be a promising method in the field of neuropsychiatry based on its unique advantages.

The gut-eye axis:from brain neurodegenerative diseases to age-related macular degeneration

Age-related macular degeneration is a serious neurodegenerative disease of the retina that significantly impacts vision.Unfortunately,the specific pathogenesis remains unclear,and effective early treatment options are consequently lacking.The microbiome is defined as a large ecosystem of microorganisms living within and coexisting with a host.The intestinal microbiome undergoes dynamic changes owing to age,diet,genetics,and other factors.Such dysregulation of the intestinal flora can disrupt the microecological balance,resulting in immunological and metabolic dysfunction in the host,and affecting the development of many diseases.In recent decades,significant evidence has indicated that the intestinal flora also influences systems outside of the digestive tract,including the brain.Indeed,several studies have demonstrated the critical role of the gut-brain axis in the development of brain neurodegenerative diseases,including Alzheimer’s disease and Parkinson’s disease.Similarly,the role of the“gut-eye axis”has been confirmed to play a role in the pathogenesis of many ocular disorders.Moreover,age-related macular degeneration and many brain neurodegenerative diseases have been shown to share several risk factors and to exhibit comparable etiologies.As such,the intestinal flora may play an important role in age-related macular degeneration.Given the above context,the present review aims to clarify the gut-brain and gut-eye connections,assess the effect of intestinal flora and metabolites on age-related macular degeneration,and identify potential diagnostic markers and therapeutic strategies.Currently,direct research on the role of intestinal flora in age-related macular degeneration is still relatively limited,while studies focusing solely on intestinal flora are insufficient to fully elucidate its functional role in age-related macular degeneration.Organ-on-a-chip technology has shown promise in clarifying the gut-eye interactions,while integrating analysis of the intestinal flora with research on metabolites through metabolomics and other techniques is crucial for understanding their potential mechanisms.

生物样品中汞的电感耦合等离子体质谱分析与干扰校正方法

电感耦合等离子体质谱法(ICP-MS)在测定生物样品中的Hg时,由于Hg元素的电离能高、电离效率低,且存在W的氧化物等多原子离子干扰,很难进行直接准确测定,加之生物样品中有机质含量高,基体复杂,也会导致分析结果产生偏差。本文通过比较标准模式(STD)和动能歧视模式(KED)下测定Hg的质谱干扰情况,表明在STD模式下200Hg、202Hg均受到W、Re、Os、Er、Dy等元素多原子离子的干扰,而KED模式有效地降低了干扰;在KED模式下选择202Hg作为分析同位素,Er、Dy、Re、Os等对Hg的干扰可以忽略不计,而W的氧化物干扰仍难以完全消除。进而详细研究了KED模式下W对Hg测定的质谱干扰,Hg所受干扰程度与W含量呈线性相关(R2=0.9997),可利用KED模式结合数学校正法消除W的质谱干扰;优选了样品稀释倍数和内标元素,选择稀释倍数为100倍,50μg/L的Rh作为内标补偿基体效应。在此基础上建立生物样品中Hg的ICP-MS分析与干扰校正方法,检出限为1.2ng/g。采用该方法对9个标准物质中Hg含量进行测定,测定值与标准值(或参考值)一致,尤其是国家标准物质GBW10028(黄芪)、GBW10025(螺旋藻)、GBW10015a(菠菜)的准确度显著提高,相对标准偏差(n=10)为0.7%~7.0%。该方法操作简便,适用于W含量范围在0~1000ng/g,Hg含量范围在3.2~670ng/g的生物样品的测试。

Mer受体酪氨酸激酶与SD大鼠糖尿病周围神经病变的相关性

背景:糖尿病周围神经病变的发病机制目前尚未明确,TAM(Tyro3、Axl和MerTK)受体酪氨酸激酶具有控制中枢神经系统凋亡细胞和抑制炎症反应的作用。目的:探讨2型糖尿病及其周围神经病变SD大鼠血浆及坐骨神经组织Mer受体酪氨酸激酶水平的差异,研究Mer受体酪氨酸激酶与糖尿病周围神经病变的关联性。方法:40只雄性SD大鼠随机分为对照组15只、2型糖尿病组10只及2型糖尿病周围神经病变组15只。对照组大鼠喂普通饲料,实验组大鼠行高脂高糖饮食6周,并腹腔内注射单剂量小剂量链脲佐菌素35 mg/kg,14 d尾静脉取血检测血糖≥16.7 mmol/L为2型糖尿病造模成功。周围神经病变组大鼠动物继续高糖、高脂喂养8周。麻醉后活体分离检测大鼠坐骨神经传导速度,腹主动脉取血,取坐骨神经组织。光镜下观察各组神经纤维组织学改变,确认糖尿病周围神经病变造模成功。ELISA检测大鼠外周血血糖、血脂及血清MerTK水平,苏木精-伊红染色观察坐骨神经组织学改变;免疫荧光、免疫组化及免疫蛋白印迹检测坐骨神经组织中MerTK的表达。结果与结论:①实验成功构建SD大鼠2型糖尿病及2型糖尿病周围神经病变大鼠模型,糖尿病周围神经病变成模率80%;与对照组比较,2型糖尿病及糖尿病周围神经病变组大鼠血糖水平显著升高(P<0.0001),糖尿病周围神经病变组高于2型糖尿病组;坐骨神经传导速度明显下降(P<0.05),糖尿病周围神经病变组低于2型糖尿病组。②组织学检查:与对照组相比,2型糖尿病组大鼠坐骨神经核减少,有部分空泡变性,出现吞噬细胞;糖尿病周围神经病变组胞体肿胀,核间距增大,出现空泡变性,髓鞘周围出现隔断、不平滑,轴索变为网格状。③血清中MerTK水平:糖尿病周围神经病变组显著高于对照组(P<0.05)。④坐骨神经组织中MerTK的表达:与对照组比较,糖尿病周围神经病变组中明显上调(P<0.05)。⑤结论:糖尿病周围神经病变大鼠血浆及坐骨神经组织中Mer受体酪氨酸激酶水平升高,推测与其抗炎及免疫调节作用有关。

Optimization of the treatment with immunosuppressants and biologics in inflammatory bowel disease

Many placebo controlled trials and meta-analyses evaluated the efficacy of different drugs for the treatment of inflammatory bowel disease (IBD), including immunosuppressants and biologics. Their use is indicated in moderate to severe disease in non responders to corticosteroids and in steroid-dependent patients, as induction and maintainance treatment. Infliximab, as well as cyclosporine, is considered a second line therapy in the case of severe ulcerative colitis, or non-responders to intravenous corticosteroids. An adequate dosage and duration of therapy with thiopurines should be reached before evaluating their efficacy. Methotrexate is a valid option in patients with Crohn&#x02019;s disease but its use is confined to patients who are intolerant or non-responders to thiopurines. Evidence for the use of methotrexate in ulcerative colitis is insufficient. The use of thalidomide and mycophenolate mofetil is not recommended in patients with inflammatory bowel disease, these treatments could be considered in case of failure of all other therapeutic options. In patients with moderately active ulcerative colitis, refractory to thiopurines, the use of tacrolimus is considered an alternative to biologics. An increase of the dose or a decrease in the interval of administration of biological treatment could be useful in the presence of an incomplete clinical response. In the case of primary failure of an anti-tumor necrosis factor alpha a switch to another one should be considered. Data on the efficacy of combination therapy are up to now insufficient to consider this strategy in all IBD patients. The final outcome of the treatment should be considered the clinical remission, with mucosa healing, and not the clinical response. The evaluation of serum concentration of thiopurine methyl transferase activity, thiopurine metabolites, biologic serum levels and antibiologic antibodies could be useful for the management of the treatment but it has not been routinely applied in clinical practice. The evidence of high risk development of lymphoma and cutaneous malignancies should be considered in patients treated with immunosuppressants and biologics for a long period.

Risk of hepatitis B virus reactivation in patients with autoimmune diseases undergoing non-tumor necrosis factor-targeted biologics

Hepatitis B virus reactivation(HBVr)can occur in patients treated with immunosuppressive medications.Risk stratification for HBVr based on hepatitis B virus(HBV)serology and viral load is an important strategy to determine appropriate HBV monitoring and antiviral prophylaxis use.Recent advances in the understanding of pathophysiology of autoimmune diseases have led the development of cytokine-targeted therapies.Tumor necrosis factor(TNF)-αinhibitors have been widely used for patients with inflammatory bowel disease,psoriasis,and rheumatic diseases.Further,the clinical benefits of interleukin(IL)-12/23,IL-17,or Janus kinases inhibitors have been demonstrated in these patients.It is well known that TNF-αinhibitor use can lead to HBVr,however,the risk of HBVr in patients undergoing non-TNF-targeted biologics have not been fully understood.In this review,we discuss the risk of HBVr in patients treated with non-TNF-targeted biologics,and immunological mechanisms of these medications causing HBVr.

Precision medicine in inflammatory bowel disease:Individualizing the use of biologics and small molecule therapies

The advent of biologics and small molecules in inflammatory bowel disease(IBD)has marked a significant turning point in the prognosis of IBD,decreasing the rates of corticosteroid dependence,hospitalizations and improving overall quality of life.The introduction of biosimilars has also increased affordability and enhanced access to these otherwise costly targeted therapies.Biologics do not yet represent a complete panacea:A subset of patients do not respond to first-line anti-tumor necrosis factor(TNF)-alpha agents or may subsequently demonstrate a secondary loss of response.Patients who fail to respond to anti-TNF agents typically have a poorer response rate to second-line biologics.It is uncertain which patient would benefit from a different sequencing of biologics or even a combination of biologic agents.The introduction of newer classes of biologics and small molecules may provide alternative therapeutic targets for patients with refractory disease.This review examines the therapeutic ceiling in current treatment strategies of IBD and the potential paradigm shifts in the future.

固态胶原基材料在医疗器械中的应用

背景:胶原蛋白是细胞外基质的主要构成成分,具有稳定的三螺旋结构特性、良好的生物学特性和可加工性,是一种理想的生物医用材料,已被广泛应用于医疗器械领域。目的:分析固态胶原基材料的主要产品形态、技术特点、制备工艺和临床应用。方法:应用计算机检索PubMed和中国知网数据库收录的有关固态胶原基材料在医疗器械领域应用的研究,英文检索词为“collagen,collagen sponge,collagen membrane,collagen powder,collagen gel,decellularized extracellular martix,medical device,clinical application”,中文检索词为“胶原,胶原海绵,胶原膜,胶原粉,胶原凝胶,脱细胞基质,医疗器械,临床应用”。根据纳入与排除标准对所有文章进行初筛后,保留质量和相关性较高的92篇文献进行综述。结果与结论:以动物源来源固态胶原基材料作为医疗器械制备材料,已在原料提取、制备工艺及临床应用等方面取得显著进展,海洋生物胶原和重组胶原增加了胶原来源的可选择性。固态胶原基医疗器械产品类型丰富,制备方法多样,其中交联改性是提升材料性能的重要途径,值得被深入研究。在产品开发思路上,固态胶原基医疗器械产品研究正朝着立体化和仿生化方向发展,与其他生物材料、药物复合的研究也展现出巨大潜力。尽管国内已有不少胶原基医疗器械应用于临床,但在人工皮肤、软骨修复和神经损伤修复等领域的产品开发仍显不足,需在技术和产业化上进一步突破与创新。

Are we giving biologics too late? The case for early versus late use

Corticosteroids and immunomodulators have been the mainstay therapies for Crohn’s disease. Corticosteroids are highly effective to control symptoms in the short- term, but they are not effective in maintaining remission, they heal the mucosa in a reduced proportion of cases, and long-time exposure is associated with an increased risk of infections and mortality. Immunomodulators, azathioprine and methotrexate, heal the mucosa in a higher proportion of patients that corticosteroids but their onset of action is slow and they benefit less than half of patients with Crohn’s disease. In the last decade, medical therapy for Crohn’s disease has experienced a remarkable change due to the introduction of biologic therapy, and particularly the use of anti-tumour necrosis factor-alpha agents. Infliximab, adalimumab, and certolizumab pegol have demonstrated efficacy for induction and maintenance of remission in active Crohn’s disease. These agents have raised the bar for what is a suitable symptomatic response in Crohn’s disease and modification of the natural history of the disease has become a major goal in the treatment of Crohn’s disease. There are several data in the literature that suggest that early use of biologic therapy and achievement of mucosal healing contribute to disease course modification. However, many questions on early biological therapy for Crohn’s disease remain still unanswered.

Role of the combination of biologics and/or small molecules in the treatment of patients with inflammatory bowel disease

Inflammatory bowel disease(IBD)is a group of chronic diseases that includes ulcerative colitis,Crohn’s disease,and indeterminate colitis.Patients with IBD require prolonged treatment and high utilization of healthcare resources for proper management.The treatment of patients with IBD is focused on achieving therapeutic goals including clinical,biochemical,and endoscopic variables that result in improvement of the quality of life and prevention of disability.Advanced IBD treatment includes tumor necrosis factor inhibitors,integrin antagonist,antagonist of the p40 subunit of interleukin 12/23,and small molecule drugs.However,despite the multiple treatments available,about 40%of patients are refractory to therapy and present with persistent symptoms that have a great impact on their quality of life,with hospitalization and surgery being necessary in many cases.Dual therapy,a strategy sometimes applicable to refractory IBD patients,includes the combination of two biologics or a biologic in combination with a small molecule drug.There are two distinct scenarios in IBD patients in which this approach can be used:(1)Refractory active luminal disease without extraintestinal manifestations;and(2)patients with IBD in remission,but with active extraintestinal manifestations or immune-mediated inflammatory diseases.This review provides a summary of the results(clinical response and remission)of different combinations of advanced drugs in patients with IBD,both in adults and in the pediatric population.In addition,the safety profile of different combinations of dual therapy is analyzed.The use of newer combinations,including recently approved treatments,the application of new biomarkers and artificial intelligence,and clinical trials to establish effectiveness during long-term followup,are needed to establish new strategies for the use of advanced treatments in patients with refractory IBD.

Is non-biological treatment of rheumatoid arthritis as good as biologics?

The management of rheumatoid arthritis(RA) in the past three decades has undergone a paradigm shift from symptomatic relief to a "treat-to-target" approach. This has been possible through use of various conventional and biologic disease modifying anti-rheumatic drugs(DMARDs) which target disease pathogenesis at a molecular level. Cost and infection risk preclude regular use of biologics in resource-constrained settings. In therecent years, evidence has emerged that combination therapy with conventional DMARDs is not inferior to biologics in the management of RA and is a feasible cost-effective option.

Current status of novel biologics and small molecule drugs in the individualized treatment of inflammatory bowel disease

Treatment strategies for inflammatory bowel disease(IBD)are rapidly evolving with the development of biologics and small molecule drugs(SMDs).However,these drugs are not guaranteed to be effective in all patients,and a“ceiling effect”of biologic monotherapy may occur.This issue highlights an unmet need for optimizing the use of biologics and predicting therapeutic responses.Thus,the development of new drugs with novel mechanisms of action is urgently needed for patients with primary nonresponse and secondary loss of response to conventional biologics and SMDs.In addition,combining different biologics or SMDs has been proposed as a novel strategy to enhance treatment efficacy in IBD,which theoretically has multidimensional anti-inflammatory potential.Based on the current evidence available for IBD,dual targeted therapy may be a promising strategy for refractory IBD patients who have failed in multiple biologic treatments or who have extraintestinal manifestation.Additionally,identifying the subgroup of IBD patients who are responding to biological combination therapies is also equally important in stable disease remission.In this review,we summarize the newly developed biologics and SMDs and the current status of biologics/SMDs to highlight the development of individualized treatment in IBD.

Effectiveness, safety, and drug sustainability of biologics in elderly patients with inflammatory bowel disease: A retrospective study

BACKGROUND Biologic therapy resulted in a significant positive impact on the management of inflammatory bowel disease(IBD) however data on the efficacy and side effects of these therapies in the elderly is scant.AIM To evaluate retrospectively the drug sustainability, effectiveness, and safety of the biologic therapies in the elderly IBD population.METHODS Consecutive elderly(≥ 60 years old) IBD patients, treated with biologics [infliximab(IFX), adalimumab(ADAL), vedolizumab(VDZ), ustekinumab(UST)] followed at the McGill University Inflammatory Bowel Diseases Center were included between January 2000 and 2020.Efficacy was measured by clinical scores at 3, 6-9 and 12-18 mo after initiation of the biologic therapy. Patients completing induction therapy were included. Adverse events(AEs) or serious AE were collected during and within three months of stopping of the biologic therapy.RESULTS We identified a total of 147 elderly patients with IBD treated with biologicals during the study period, including 109 with Crohn’s disease and 38 with ulcerative colitis. Patients received the following biologicals: IFX(28.5%), ADAL(38.7%), VDZ(15.6%), UST(17%). The mean duration of biologic treatment was 157.5(SD = 148) wk. Parallel steroid therapy was given in 34% at baseline,19% at 3 mo, 16.3% at 6-9 mo and 6.5% at 12-18 mo. The remission rates at 3, 6-9 and 12-18 mo were not significantly different among biological therapies. Kaplan-Meyer analysis did not show statistical difference for drug sustainability(P = 0.195), time to adverse event(P = 0.158) or infection rates(P = 0.973) between the four biologics studied. The most common AEs that led to drug discontinuation were loss of response, infusion/injection reaction and infection.CONCLUSION Current biologics were not different regarding drug sustainability, effectiveness, and safety in the elderly IBD population. Therefore, we are not able to suggest a preferred sequencing order among biologicals.

Predictive value of blood concentration of biologics on endoscopic inactivity in inflammatory bowel disease:A systematic review

BACKGROUND Although blood concentration of biologics is an important composition of disease management in inflammatory bowel disease(IBD)patients,complexity and uncertainty of biological management encourage many disputes in predicting the outcome of IBD patients through blood concentration of biologics.AIM To verify the predictive value of blood concentration of biologics on endoscopic inactivity in IBD patients under different situations.METHODS We searched PubMed/MEDLINE,Embase,and Web of Science up to May 2020 and identified IBD patients as the research cohort as well as the correlations between blood concentration of biologics and endoscopic inactivity in IBD patients as the research direction.RESULTS A total of 23 articles with 30 clinical studies and 1939 IBD patients were included.The predictive cut-off value of blood concentration of infliximab on mucosal healing should be 2.7-10.6μg/mL in IBD.Blood concentration of infliximab reaching 5.0-12.7μg/mL or more increased the probability of fistula healing/closure in perianal fistulizing Crohn's disease.Blood concentration of adalimumab reaching 7.2-16.2μg/mL or more could predict mucosal healing in IBD.The predictive cut-off value of blood concentration of adalimumab on fistula healing/closure should be 5.9-9.8μg/mL in perianal fistulizing Crohn's disease.Blood concentration of vedolizumab surpassing 25.0μg/mL indicated mucosal healing in ulcerative colitis patients under maintenance therapy and the predictive cut-off value of blood concentration on mucosal healing or endoscopic remission under induction therapy in IBD could be 8.0-28.9μg/mL.CONCLUSION Blood concentration of biologics should not be utilized to predict endoscopic inactivity of IBD independently due to discrepancies in clinical studies,whereas conducting therapeutic drug monitoring intensively contributes to precise therapy.

Are we giving biologics too much time? When should we stop treatment?

The optimal duration of biological treatment, particularly anti-TNF, in inflammatory bowel disease （IBD） is a very important question both for patients and physicians. There is no published evidence to clearly and definitely answer this question. However data on natural history of IBD, long term safety of biologics, immunosuppressors （IS） cessation and some preliminary studies on biologics cessation may help us to discuss this topic. The decision to stop a biological treatment is currently based on a compromise between the benefits and risks associated with the prolongation of this treatment. IBD, more particularly CD, are characterized by the development of complications and the need for recurrent hospitalizations and surgeries in approximately 2/3 of cases. In these patients potentially in need of biological treatments, it is probable that, as it has been demonstrated for IS, the longer a stable remission has be achieved under treatment, the lower the risk of relapse is alter treatment cessation. Further prospective studies should now aim at disclosing patient characteristics associated with a low risk of relapse to imple- ment this strategy.

PLGA/赖氨酸接枝氧化石墨烯纳米粒子复合支架对MC3T3细胞成骨分化的影响

背景:骨损伤后如何有效促进骨再生和骨重建一直是临床骨修复研究中的关键问题,利用生物和降解材料搭载生物活性因子在骨修复中具有优秀的应用前景。目的:探究赖氨酸接枝氧化石墨烯(LGA-g-GO)纳米粒子改性的聚乳酸-羟基乙酸共聚物(PLGA)复合支架对成骨分化及新骨形成的影响。方法:将PLGA溶于二氯甲烷中,利用溶剂挥发法制备PLGA支架;将氧化石墨烯均匀分散于PLGA溶液中制备PLGA/GO复合支架;利用化学接枝法制备LGA-g-GO纳米粒子,将LGA-g-GO纳米粒子按不同的质量比(1%,2%,3%)与PLGA共混构建出PLGA/LGA-g-GO复合支架。表征5组支架的微观形貌、亲水性、蛋白吸附能力。将MC3T3细胞分别接种于5组支架表面,检测细胞增殖与成骨分化情况。结果与结论:①扫描电镜下可见PLGA支架表面光滑平坦,其余4组支架表面粗糙,并且随着LGA-g-GO纳米粒子加入量的增加,复合支架的表面粗糙程度加重;PLGA/LGA-g-GO(3%)复合支架的水接触角低于其他4组支架(P<0.05);PLGA/LGA-g-GO(1%,2%,3%)复合支架的蛋白吸附能力强于PLGA、PLGA/GO支架(P<0.05);②CCK-8检测显示,PLGA/LGA-g-GO(2%,3%)复合支架可促进MC3T3细胞的增殖;碱性磷酸酶染色与茜素红染色显示,PLGA/LGA-g-GO(2%,3%)组细胞碱性磷酸酶活性高于其他3组(P<0.05),PLGA/GO组、PLGA/LGA-g-GO(1%,2%,3%)组钙沉积多于PLGA组(P<0.05);③结果表明,PLGA/LGA-g-GO复合支架能够促进成骨细胞的增殖和成骨分化,有利于骨损伤后的骨再生和骨重建。

生物3D打印仿生支架促进肩袖损伤后的愈合

背景:大多数肩袖损伤发生于冈上肌腱,由于肌腱组织缺少血管以及肩袖复杂的解剖结构,临床治疗效果非常有限。组织工程技术和干细胞生物学的快速发展为提高肌腱修复质量带来了新的希望。目的:通过生物3D打印技术制备人脐带间充质干细胞/甲基丙烯酸酐化明胶复合支架,观察该支架修复肩袖损伤的效果。方法:(1)体外细胞实验:制备明胶微载体,将人脐带间充质干细胞接种于明胶微载体表面构建组织工程化干细胞。制备甲基丙烯酸酐化明胶水凝胶打印墨水,使用甲基丙烯酸酐化明胶水凝胶打印墨水重悬组织工程化干细胞,放入3D打印机的生物墨水容器中进行打印,蓝光照射固化5 min后即得人脐带间充质干细胞/甲基丙烯酸酐化明胶复合支架。通过死活染色、CCK-8实验检测支架内人脐带间充质干细胞的活性。(2)动物体内实验:采用随机区组设计方法将24只SD大鼠随机4组,每组6只:正常组不进行任何处理,肩袖损伤组、单纯支架组、细胞支架组建立冈上肌腱撕裂的肩袖损伤模型,单纯支架组、细胞支架组造模后分别将甲基丙烯酸酐化明胶支架、人脐带间充质干细胞/甲基丙烯酸酐化明胶复合支架植入肌腱损伤处。术后4周,分别进行大鼠行为学测试及肩袖冈上肌腱组织学病理组织形态观察。结果与结论:(1)体外细胞实验:死活染色结果显示,明胶微载体可减轻3D打印过程对人脐带间充质干细胞造成的损伤,随着培养时间的延长,支架内的人脐带间充质干细胞存活率升高;CCK-8实验结果显示,随着培养时间的延长,支架内的人脐带间充质干细胞活性无明显变化。(2)动物体内实验:行为学测试结果显示,相较于肩袖损伤组、单纯支架组,细胞支架组大鼠四肢运动功能明显改善;肩袖冈上肌腱苏木精-伊红与Masson染色结果显示,相较于肩袖损伤组、单纯支架组,细胞支架组肌纤维排列较规律,无明显的炎性细胞浸润,胶原容积百分比降低;免疫荧光染色结果显示,相较于肩袖损伤组、单纯支架组,细胞支架组肩袖冈上肌腱内白细胞介素6、肿瘤坏死因子α蛋白表达明显降低。(3)结果表明,生物3D打印的人脐带间充质干细胞/甲基丙烯酸酐化明胶复合支架可促进肩袖损伤组织修复再生。

重组慢病毒转染兔骨髓间充质干细胞与脱钙骨基质构建转基因组织工程材料

背景:缺损组织的修复重建受限于自体或异体可替代移植材料的来源问题而导致临床应用受限,转基因干细胞和组织工程材料研究开辟了新的治疗思路。目的:探究增强型绿色荧光蛋白重组慢病毒转染兔骨髓间充质干细胞在体外的生物学特性以及与脱钙骨基质体外构建转基因组织工程材料的生物矿化特性。方法:细胞贴壁及密度离心法获得兔骨髓间充质干细胞,增强型绿色荧光蛋白重组慢病毒以感染复数为100转染第5代兔骨髓间充质干细胞,体外观察转染细胞与未转染细胞的增殖能力、细胞表型、细胞周期以及成骨诱导后碱性磷酸酶、Runx2、骨钙素表达的差异;增强型绿色荧光蛋白重组慢病毒转染骨髓间充质干细胞与脱钙骨基质在体外构建转基因组织工程材料,对其进行扫描电镜观察及元素能谱分析。结果与结论:增强型绿色荧光蛋白重组慢病毒成功转染骨髓间充质干细胞后,在转染24,48 h细胞增殖较未转染细胞缓慢(P <0.05);在转染72 h后,细胞表型未发生变异,细胞周期、细胞增殖能力以及成骨诱导后碱性磷酸酶、Runx2、骨钙素表达量与未转染细胞无明显差异(P> 0.05);增强型绿色荧光蛋白标记的骨髓间充质干细胞在脱钙骨基质支架上有较好的生物相容性,根据荧光表达强度推测目的基因在2周左右发挥最大生物学功能,且出现了钙磷矿化物沉积,体现出优越的生物矿化特性。

Biologics in non-infectious uveitis past,present and future

Our increase in knowledge of the pathophysiology of non-infectious uveitis(NIU)and other immune-mediated diseases has been mirrored over the last two decades by the expansion of therapeutic options in the realm of immunosuppressive medications.Principal among these advances is the emergence of biologics,which offer the promise of targeted therapy and the hope of reduced toxicity when compared to corticosteroids and“standard”immunosuppression.Among the biologics,monoclonal antibodies blocking tumor necrosis factor alpha(TNF-α)have been shown to be a very effective therapeutic target for uveitis and many associated systemic inflammatory diseases.Multiple TNF blockers have shown benefit for uveitis,and in 2016,adalimumab became the first biologic and non-corticosteroid immunosuppressive to obtain Food and Drug Administration(FDA)approval in the treatment of NIU.Although effective,TNF blockers are not universally so,and safety concerns such as infection and demyelinating disease must be carefully considered and ruled out prior to their use,especially in patients with intermediate uveitis with which multiple sclerosis is a known association.Ongoing study has identified novel targets for regulation in the treatment of immune-mediated and inflammatory diseases.Interferons,interleukin and Janus kinase inhibitors in addition to antibodies targeting T cell and B cell activation highlight the expanding field of treatment modalities in NIU.Ongoing study will be required to better determine the safety and efficacy of biologics in the armamentarium of immunosuppressive treatments for NIU.