Metabolomics comparison of cord and peripheral blood-derived serum eye drops for the treatment of dry eye disease

Allogeneic peripheral blood-derived (PBS) serum eye drops have been largely used in the treatment of dry eye disease (DED). Recently, cord blood has emerged as an effective alternative serum source (cord blood serum, CBS), containing a higher amount of growth factors than PBS, it holds the promise of a better capability to stimulate corneal healing. However, the lack of a standardized method for preparation, dispensation, storage and a poor biochemical characterization still hamper the establishment of a clinical consensus. Here the metabolomes of the two different serum eye drop preparations were compared using proton nuclear magnetic resonance spectroscopy. We found that both PBS and CBS contained several organic compounds, the majority of them already detected in human tears and may be thereby considered lacrimal substitutes. Metabolites having in the multivariate statistical analysis Partial least squares discriminant analysis (PLS-DA) a VIP scores > 1.0 were considered to be significantly different. All the metabolites identified were found to have a p < 0.05 in the univariate analysis. CBS, in particular, showed the highest amount of choline, myo-inositol, glutamine, creatine and β-hydroxybutyrate. These evidences constitute relevant advances towards serum eye drops characterization and confirm that cord blood is a valid alternative source of serum eye drops.     

Blood derived eye drops for the treatment of cornea and ocular surface diseases

The use of blood derived eye drops for the treatment of ocular surface disorders has become increasingly popular in recent years. The mechanism of action is the stimulation of cellular proliferation and migration by supplying an active mixture of growth factors and cytokines at the ocular surface, thus mimicking the function of the lacking natural tears. Blood derived eye drops have been used in the last decades for the treatment of a variety of ocular surface diseases, including mainly dry eye disease, persistent corneal epithelial defect, corneal ulcer, ocular surface burn, recurrent corneal erosion and limbal stem-cell deficiency. Among overall blood derived eye drops, both autologous (from the patients themselves) and homologous (from donors) products exist, with different advantages and disadvantages. Autologous serum, obtained from the patient's own peripheral blood, is the first introduced and most commonly used product. Despite several randomized clinical trials showed its safety and efficacy, a recent Cochraine meta-analysis failed to show significant results due to low evidence. Homologous sources including allogeneic serum obtained from healthy donors, and umbilical cord blood serum collected at the time of delivery, are efficient alternatives, especially when autologous serum therapy is contraindicated or not appropriate. Platelet-derived eye drops are prepared and used in various but poor standardized preparations, namely platelet-rich plasma, plasma rich in growth factors, and platelet lysate. Future perspectives of blood-derived products include the introduction of tailored eye drops, screened for the proper content of growth factors and cytokines according to each patient and ocular surface disease.     

Low-cost protocol for the production of autologous serum eye drops by blood collection and processing centres for the treatment of ocular surface diseases

Objectives: To create a low‐cost protocol for the production of autologous serum eye drops (ASEs) by blood collection and processing centres using standard equipment and staff already available. The protocol must observe local and federal regulatory authorities' requirements for good manufacturing practice. Aim: The final aim is to improve accessibility to ASEs for patients who need them and to implement similar production parameters at all institutions producing ASEs. Background: Despite evidence of the beneficial effects of ASEs in the treatment of severe ocular surface diseases, ASEs are not approved by most federal regulatory bodies in the United States or Europe. In some countries, such as the United States, access to ASEs is extremely limited and cost‐prohibitive. Blood processing centres are in optimal position to help in increasing accessibility and lowering the cost of these products. Materials/Methods: Standard blood processing equipment, staff and materials were chosen for the protocol. The protocol was designed to minimise the risk of contamination as much as possible. Results: A low‐cost, open system protocol for production of ASEs was generated that can be implemented by most blood collection and processing centres in the United States and Europe. Conclusion: Efforts should be made to propagate a similar protocol for the production of ASEs in blood centres capable of collecting and processing blood products, making this service affordable and uniformly accessible to patients.     

新生儿脐血与外周血早期血糖监测190例分析

目的探讨新生儿脐血血糖与新生儿外周血（足底血）血糖之间的关联性,为临床早期干预提供依据。方法选择100例妊娠期无并发症及合并症的正常妊娠分娩且新生儿Apgar评分≥8分的新生儿;90例“高危儿”病例作为对照,其中妊高征30例,早产儿30例,糖尿病30例。分娩时即时抽取脐静脉血送化验室查血糖、糖化血红蛋白,同时取脐血及新生儿足底血查快速血糖值。结果快速血糖值与化验室报告血糖值有一致性,脐血血糖与外周血血糖具有显著正相关性。正常妊娠分娩的新生儿中有5％比率的低血糖。结论为避免忽略正常新生儿中的低血糖患者,可用无创的脐血查快速血糖代替新生儿足底血血糖的测定,以减少对新生儿的损伤。     

脐血输注对妇科恶性肿瘤患者外周血中IL-2、IL-6和sIL-2R的诱导表达和T细胞亚群改变

目的　探讨妇科恶性肿瘤患者免疫功能的变化机制及脐血输注对其的影响。方法　将化疗后妇科恶性肿瘤患者 30例分为脐血治疗组 15例、非脐血治疗组 15例 ,并设正常人对照组 30例 ,采用MTT法、ELISA法和流式细胞仪检测的方法观察脐血输注后患者免疫功能变化。结果　①患者外周血CD4 + 细胞含量低于正常人、CD8+细胞含量高于正常人 (t均为 2 .5 6 ,P <0 .0 5 ) ,CD4 + /CD8+ 比例倒置 ;脐血治疗组 30名脐血输注后患者CD3+ 细胞含量明显升高 (t=3.0 1,P <0 .0 1)、CD4 + 细胞含量明显升高 (t=2 .74 ,P <0 .0 5 ) ,CD4 /CD8比例恢复正常。②30名患者外周血IL 2活性较正常人降低 ,可溶性IL 2R(sIL 2R)含量明显增加 (t分别为 4 .33、5 .89,P <0 .0 1) ;脐血输注后患者IL 2活性增加 ,sIL 2R含量降低 ,与对照组相比 ,差异有显著性 (t分别为 2 .32、2 .4 2 ,P <0 .0 5 )。而IL 6活性有一定程度下降 ,但无统计学意义 (t=0 .2 7,P >0 .0 5 )。结论　妇科恶性肿瘤患者的免疫功能低于正常人 ,脐血输注后可以通过免疫活性细胞与淋巴因子水平的变化调节患者免疫功能。     

比较脐血与成人外周血中T淋巴细胞亚群分布及其因子产生的差异

目的：探讨脐血与成人外周血中淋巴细胞亚群分布及细胞因子的差异及其与移物抗宿主病（GVHD）发生的关系,方法：采用流式细胞术,三色荧光单克隆抗体标记对脐血和成人外周血中淋巴细胞进行分类,并对其分泌的IL－2,TNF－α和IFN－r进行分析比较,结果：（1）脐血中T淋巴细胞以CD^4 ,CD45RA^ 细胞为主,而成人外周血以CD^ ,CD4R5RO^ 和CD^ 8CD450^ 两种表型为主,（2）脐血淋巴细胞产生IL－2,TNF－α和IFN－r的量明显低于成人外周血淋巴细胞,进一步分析表明,脐血中产生细胞因子的细胞大部分为CD^ 4CD45^ 细胞,而成人外周血中大部分为CD^ 4CD45RO^ 和CD^ 8CD^ 45RO细胞。结论：成人外周血与脐血淋巴细胞不仅存在亚群比例上的差异,而且亚群分泌细胞因子的量也明显减少,脐血干细胞移植的GVHD发生率较低可能与脐血中淋巴细胞亚群的比例不同及分泌细胞因子减少有关。     

角膜缘干细胞移植术后应用玻璃酸钠滴眼液对翼状胬肉复发率及眼表症状的影响

目的 探讨角膜缘干细胞移植术(LCAT)后应用玻璃酸钠滴眼液预防翼状胬肉的效果及对眼表症状的影响.方法 选择2018年11月至2019年11月住院治疗的66例翼状胬肉患者,以随机抽签法将其分为观察组和对照组,每组33例.所有患者均行翼状胬肉切除术配合LCAT,对照组术后予以抗生素联合糖皮质激素滴眼,观察组在对照组基础上...     

Investigating the production of platelet lysate obtained from low volume Cord Blood Units: Focus on growth factor content and regenerative potential

BackgroundThe regenerative potential of platelet lysate (PL) and platelet gel (PG) is mediated by the release of platelets (PLTs) growth factors. The aim of this study was the evaluation of the PL production utilizing low volume single Cord Blood Units (CBUs) and the comparison of the biomolecule content between PLs obtained from intermediate and high volume CBUs.MethodsCBUs (n = 90) with volumes greater than 50 ml and initial platelet count > 150 × 10⁹/L were used. CBUs were classified into the following groups: group A (50–80 ml), group B (81–110 ml) and group C (111–150 ml). The CBUs were centrifuged twice for the production of the platelet concentrate (PC), which was stored at ? 80 °C for at least 48 h. Then, rapidly thawed and the biomolecule content was determined using commercial ELISA kits. The regenerative potential of PLs was evaluated using the scratch wound and in vitro angiogenesis assay.ResultsCBPL was produced from low volume single CBUs and contained 3.4 ± 0.3 ×10⁹ PLTs. PL obtained from intermediate and high volume CBUs consisted of 10.2 ± 0.3 and 16.1 ± 0.4 × 10⁹ PLTs. All PL groups were characterized by high biomolecule content. Gap closure was observed within 72 h after the wound assay initiation and the capillary tubes were formed in all study groups.ConclusionThis study provided significant evidence regarding the utilization of the low volume CBUs for the production of CB derivatives, thus can serve as healing mediators in regenerative medicine approaches.     

Topical treatment of ocular surface defects: comparison of the epitheliotrophic capacity of fresh frozen plasma and serum on corneal epithelial cells in an in vitro cell culture model

Accelerated healing of ocular surface disorders was reported using serum for topical application. It is supposed that growth factors, fibronectin and vitamins in serum support the proliferation of corneal epithelial cells. The use of fresh frozen plasma (FFP) instead of serum is theoretically attractive, as it is more easily available from blood banks. In this study, serum and FFP were investigated for composition of epitheliotrophic factors and effect on corneal epithelial cells. Whole blood was taken from five donors. Serum and FFP were prepared, and the concentrations of epithelial growth factor (EGF), Platelet-derived growth factor (PDGF), transforming growth factor-beta1, fibronectin and vitamin A were determined. Immortalized human corneal epithelial cells were used to investigate growth, migration and differentiation in response to both blood products. Significant differences were found regarding the mediator composition of serum and FFP. Serum rather than FFP was significantly superior in stimulating cell growth, migration and differentiation. The epitheliotrophic capacity of blood products depends upon the composition of growth factors and vitamins. Blood clotting strongly influences the growth factor pattern. The superior epitheliotrophic capacity of serum might be due to the higher concentration of proliferation mediators such as EGF and PDGF and its higher content of vitamin A.     

巨大儿、宫内发育迟缓儿的脐血胃泌素、生长激素水平变化

目的研究巨大儿、宫内发育迟缓(IUGR)患儿的发生与脐血中胃泌素(Gas)、生长激素(GH)的关系,以探讨内分泌环境对胎儿生长发育的影响。方法检测正常儿组(71名)、IUGR组(31例)和巨大儿组(28例)的脐血Gas、GH水平。结果正常儿组中,经择期剖宫产的研究对象与经阴式分娩的相比,脐血Gas水平显著降低(P<0.01),脐血GH水平明显升高(P<0.05);采用阴式分娩的研究对象中,IUGR患儿脐血GH水平显著高于正常儿,巨大儿脐血Gas水平显著低于正常儿。结论胎儿体重和分娩方式对脐血Gas和GH水平有影响。     

巨大儿、宫内发育迟缓儿的脐血胃泌素、生长激素水平变化

目的 研究巨大儿、宫内发育迟缓（IUGR）患儿的发生与脐血中胃泌素（Gas）、生长激素（GH）的关系,以探讨内分泌环境对胎儿生长发育的影响.方法 检测正常儿组（71名）、IUGR组（31例）和巨大儿组（28例）的脐血Gas、GH水平.结果 正常儿组中,经择期剖宫产的研究对象与经阴式分娩的相比,脐血Gas水平显著降低（P〈0.01）,脐血GH水平明显升高（P〈0.05）;采用阴式分娩的研究对象中,IUGR患儿脐血GH水平显著高于正常儿,巨大儿脐血Gas水平显著低于正常儿.结论 胎儿体重和分娩方式对脐血Gas和GH水平有影响.     

脐血移植治疗遗传性疾病的研究进展与展望

目前,造血干细胞移植(HSCT)是唯一能治愈遗传性疾病的方法,而脐血作为造血干细胞的新来源,脐血移植(CBT)已被广泛应用于遗传性疾病的临床治疗.笔者回顾了CBT应用于多种类型遗传性疾病,如血红蛋白病、遗传代谢病、先天性免疫缺陷病(PID)及先天性骨髓衰竭性疾病的疗效,为遗传性疾病患者治疗方案的选择提供了理论依据.     

The influence of growth hormone substitution therapy on erythroid and myeloid progenitor cells and on peripheral blood cells in adult patients with growth hormone deficiency

It has been reported that hypophysectomized rats exhibit normochromic, normocytic anaemia. Pancytopenia with impaired DNA synthesis in the bone marrow can be restored in these hypophysectomized rats by syngeneic pituitary grafts placed under the kidney capsule or treatment with growth hormone (GH). Until now, adults with hypopituitarism have received adequate replacement therapy with thyroxine, cortisol and sex steroids, but not with GH. We therefore investigated the effects of GH replacement therapy on the proliferation and differentiation of erythroid and myeloid progenitor and peripheral blood cells in 11 adult patients with growth hormone deficiency in a double-blind, placebo-controlled study for the first 6 months of therapy. The placebo group showed no changes during the first 6 months without therapy in either insulin-like growth factor I (IGF-I) levels, erythroid and myeloid progenitor precursor cells or peripheral blood cells. After commencement of GH therapy, IGF-I levels rose significantly during 24 months of therapy from 75.3±13.5 to 225±34.7ngmL^?1 (P<0.001). Erythroid and myeloid progenitor precursor cells showed a steep and significant increase after 18 and 24 months of therapy (erythroid: from 10.7±3.5 to 261.4±79.8, P<0.02, after 18 months and to 276.8±149.8 × 10⁵ mononuclear cell colonies, P<0.03, after 24 months; granulocyte–macrophage colony-forming units: from 39.7±9.8 to 316.9±124.6, P<0.002, after 18 months and to 366±188.7×10⁵ mononuclear cell colonies, P<0.03, after 24 months), whereas the peripheral red and white blood cells exhibited only minimal non-significant changes. The principal regulators of erythropoiesis, such as erythropoietin, and parameters reflecting erythropoiesis in the peripheral blood, such as reticulocytes, remained almost unchanged throughout the whole study period. We therefore conclude that patients with GH deficiency do not have anaemia, but have haematopoietic precursor cells in the lower normal range, and that GH substitution therapy over a period of 24 months has a marked effect on erythroid and myeloid progenitor precursor cells but only negligible effects on peripheral blood cells in GH-deficient adults.     

脐血HLA-Ⅰ类抗原血清学分型与基因分型的比较研究

目的　探讨血清学分型方法在脐血HLA分型中的应用价值。方法　对 180 0份脐血的HLA AB分型结果进行分析 ,并与其中随机抽取的 36份标本的聚合酶链反应 序列特异性引物 (PCR SSP)分型结果进行比较分析。血清学分型分别以T免疫磁珠和淋巴细胞分层液分离细胞。结果　T免疫磁珠分离的T淋巴细胞比淋巴细胞分层液分离的单个核细胞活性更好 ;体外搁置时间较长 (2 4～48h)的脐血须用T免疫磁珠分离细胞 ;与外周血单个核细胞相比 ,脐血细胞的血清学反应强度偏低。除 2 8份因细胞分离后量少或活性低外 ,1772份脐血标本的HLA AB血清学分型中 17.5 % (310份 )有不能确定的抗原 ,其主要原因依次为交叉反应及部分分型试剂特异性不高、部分抗原反应弱、细胞活性低所致的高本底及非特异反应。 36份标本PCR SSP分型均获成功 ,与之对照 ,血清学分型有 16 .8%的错误率 ,13.9%分辨率低于PCR SSP分型。结论　血清学分型方法虽快速简便 ,但有一定的误差 ,用基因分型作为补充是必要的。     

重组人表皮生长因子凝胶联合玻璃酸钠滴眼液治疗对白内障超声乳化术后干眼症患者泪膜稳定性、泪液基础分泌量的影响

目的分析重组人表皮生长因子(rhEGF)凝胶联合玻璃酸钠滴眼液治疗对白内障超声乳化术后干眼症患者泪膜稳定性、泪液基础分泌量的影响。方法选取82例白内障术后干眼症患者,根据术后治疗方式的差异分为联合组42例和单一组40例,联合组采用rhEGF凝胶联合玻璃酸钠滴眼液治疗,单一组采用玻璃酸钠滴眼液治疗。对比两组临床疗效,治疗前及治疗2、4周的干眼症状评分。两组均于治疗前及治疗1、3月后进行泪膜厚度测量、基础泪液分泌试验(SIt)、角膜荧光素染色评分(FL)及泪膜破裂时间(BUT)检测,根据检测结果评估两组内膜结构和稳定性,同时比较治疗前及治疗1、3月后白介素-6(IL-6)、IL-1β及肿瘤坏死因子-α(TNF-α)水平。结果治疗2、4周后两组干眼症状评分较治疗前降低,且联合组评分低于单一组(P<0.05)。治疗1、3月后两组泪膜厚度、SIt及BUT升高,FL降低,观察组泪膜厚度、SIt及BUT均高于对照组,FL低于对照组(P<0.05)。治疗1、3月后两组IL-6、IL-1β及TNF-α水平下降,联合组IL-6、IL-1β及TNF-α水平均低于单一组(P<0.05)。联合组临床总有效率高于单一组(P<0.05)。两组不良反应发生率比较,差异无统计学意义(P>0.05)。结论重组rhEFG凝胶联合玻璃酸钠滴眼液可以有效改善白内障超声乳化术后干眼症患者临床症状、泪膜稳定性及眼液基础分泌量,临床疗效较仅玻璃酸钠滴眼液治疗佳,安全性高,值得临床推广。     

脐血干细胞移植及电针治疗脊髓损伤大鼠神经生长因子及神经营养因子3的表达

背景：研究表明,脐血干细胞移植对脊髓损伤的恢复起促进作用,而电针也能够通过抑制星形胶质细胞增生,来减少损伤部瘢痕形成,故推测两者结合可能在急性脊髓损伤治疗中发挥重要作用。目的：观察人脐血干细胞局部移植联合督脉电针治疗后大鼠脊髓损伤组织神经生长因子、神经营养因子3的表达。方法：选取雌性SD大鼠72只,随机分为对照组、损伤组、移植组、联合组。对照组单纯性背部切口后缝合,损伤组脊髓横断处(T 10结果与结论：脊髓损伤后,移植组与损伤组相比,联合组与移植组相比,神经生长因子、神经营养因子3在7,14,28 d表达量均增加(P<0.05)。Western Blot、实时荧光定量PCR与免疫组化结果相一致。结果显示人脐血干细胞移植与电针联合治疗脊髓损伤具有协同作用,显著上调损伤脊髓神经生长因子、神经营养因子3的表达水平,有利于脊髓损伤后功能恢复。水平)放置约1 mm×2 mm×2 mm大小、浸润生理盐水的明胶海绵;移植组及联合组在脊髓横断处放置浸润人脐血干细胞悬液的明胶海绵,联合组于造模后1 h开始给予督脉电针治疗。在相应处理7,14,28 d后应用免疫组织化学、Western Blot及实时荧光定量PCR方法检测脊髓组织神经生长因子、神经营养因子3表达量的变化。     

非血缘脐血移植治疗成年人恶性血液病的展望

近年来,非血缘脐血移植(UCBT)治疗成年人恶性血液病取得较多进展.UCBT治疗急性白血病的疗效与单倍体移植、非血缘骨髓移植、外周血干细胞移植的疗效相当,成为治疗恶性血液病的重要选择.单份UCBT的优势在于移植物抗宿主病(GVHD)发生率低且易于控制,保留有较好的移植物抗肿瘤(GVT)效应,并且患者接受治疗后生活质量较好.而UCBT的劣势在于其脐血干细胞数量少(尤其对于成年患者),因此其植入率低、植入较慢、免疫重建慢、移植早期感染发生风险高.改善UCBT的植入率低、免疫重建慢等问题,可以提高UCBT治疗成年人恶性血液病的疗效,丰富造血干细胞移植的供源.本文对今后UCBT治疗成年人恶性血液病,主要在解决UCBT植入问题、免疫重建问题、脐血人类白细胞抗原(HLA)配型等方面进行展望.     

乳腺癌组织中原癌基因BMI-1及人表皮生长因子受体-2表达与外周血微转移的关系

目的 观察乳腺癌患者乳腺癌组织中原癌基因BMI-1、人表皮生长因子受体-2(human epidermal growth factor receptor 2,HER-2)阳性表达情况,探讨乳腺癌组织BMI-1、HER-2与外周血微转移的关系.方法 89例乳腺癌患者,均采用实时荧光定量PCR法检测人乳腺珠蛋白mRNA相...     

外周血TSGF及hMAM表达指标监测乳腺癌微转移价值初探

目的探索TSGF和hMAM表达指标对乳腺癌细胞外周血微转移诊断的应用价值。方法分离受检者外周血单个核细胞,TRIZOL液提取总RNA,作RT-PCR并用-βactin监控;确认的cDNA先作普通PCR扩增,再作荧光定量PCR扩增。血清作TSGF检测。结果64例标本有63例确认转录为cDNA,其中乳腺癌43例、良性乳腺肿瘤和正常对照各10例。经普通PCR后再作荧光定量PCR,乳腺癌hMAM表达阳性率为53.3%、良性乳腺肿瘤和正常对照均无阳性,乳腺癌组与良性乳腺肿瘤或正常对照组间hMAM表达有显著差异;淋巴结转移组与未转移组,hMAM表达有显著差异。TSGF阳性率为1.6%,乳腺癌组与良性乳腺肿瘤组及正常对照组间无显著差异。结论二次PCR的灵敏度可成功检出外周血hMAM的表达,而单次PCR法的灵敏度不够;外周血hMAM表达指标可用于监测乳腺癌细胞微转移,但未见TSGF指标对乳腺癌微转移的监测价值。