Selection of oocytes for in vitro maturation by brilliant cresyl blue staining： a study using the mouse model

Selecting oocytes that are most likely to develop is crucial for in vitro fertilization and animal cloning. Brilliant cresyl blue （BCB） staining has been used for oocyte selection in large animals, but its wider utility needs further evaluation. Mouse oocytes were divided into those stained （BCB＋） and those unstained （BCB-） according to their ooplasm BCB coloration. Chromatin configurations, cumulus cell apoptosis, cytoplasmic maturity and developmental competence were compared between the BCB＋ and BCB- oocytes. The effects of oocyte diameter, sexual maturity and gonadotropin stimulation on the competence of BCB＋ oocytes were also analyzed. In the large- and medium-size groups, BCB＋ oocytes were larger and showed more surrounded nucleoli （SN） chromatin configurations and higher frequencies of early atresia, and they also gained better cytoplasmic maturity （determined as the intracellular GSH level and pattern of mitochondrial distribution） and higher developmental potential after in vitro maturation （IVM） than the BCB-oocytes. Adult mice produced more BCB＋ oocytes with higher competence than the prepubertal mice when not primed with PMSG. PMSG priming increased both proportion and developmental potency of BCB＋ oocytes. The BCB＋ oocytes in the large-size group showed more SN chromatin configurations, better cytoplasmic maturity and higher developmental potential than their counterparts in the medium-size group. It is concluded that BCB staining can be used as an efficient method for oocyte selection, but that the competence of the BCB＋ oocytes may vary with oocyte diameter, animal sexual maturity and gonadotropin stimulation. Taken together, the series of criteria described here would allow for better choices in selecting oocytes for better development.

Application of Prussian blue staining in the diagnosis of ocular siderosis

AIM:To explore the value of Prussian blue staining in the diagnosis of ocular siderosis.METHODS:Between January 2012 and January 2013,the Prussian blue stain used in anterior lens capsule and vitreous liquid after centrifugation from patients with definitive diagnosis and suspicious diagnosed of ocular siderosis. At the same time, give a negative control.RESULTS:Anterior lens capsule membrane and liquid of vitreous cavity from patients with definitive diagnosis and suspicious diagnosed of ocular siderosis revealed ferric ions that stained positively with Prussian blue. In the control group, there is no positive reaction.CONCLUSION:Prussian blue staining in the diagnosis of ocular siderosis has a very significant worth,suspected cases can be definitive diagnosed.

Application of Luxol Fast Blue staining in locating the corticospinal tract in adult rats

BACKGROUND: There are many methods for myelin staining, mordant, or oil-soluble dye or the special reaction of osmic acid with lipoid is used according to different principles. The commonly used methods are classic Well staining, classic lithium carbonate-haematine staining, fast green staining, silver staining, etc. Luxol Fast Blue can brightly stain myelin sheath, and has certain specificity. The background can be very clean if there is proper differentiation, whereas Luxol Fast Blue is cheap and convenient to operate, thus it is an ideal staining reagent for routine myelin sheath. OBJECTIVE: To show the corticospinal tract of normal adult rats with Luxol Fast Blue staining method. DESIGN: A repetitive measurement design. SETTINGS: Institute of Nuerobiology, Nantong University; Department of Rehabilitation Medicine, Affiliated Hospital of Nantong University. MATERIALS: Six healthy adult male SD rats of clean degree, weighing averagely 300 g, were provided by the experimental animal center of Nantong University. 1 g/L Luxol Fast Blue solution was provided by Sigma Company; Leica CM1900 cryostat microtome by Leica Company; Leica DMR microscope by Leica Company. METHODS: The experiment was carried out in the Staff Room of Human Anatomy, Nantong University in May 2005. The rats were given intraperitoneal injection of combined anesthetic (2 mL/kg), then the chest was open for perfusing saline and phosphate buffer containing formamint via heart. Brain and spinal cord were removed after 1 hour then fixed, then changed to phosphate buffer (pH 7.4) containing 300 g/L saccharu at 4 ℃, and stayed overnight, tissue blocks at pyramid, decussation of pyramid and cervical, thoracic, lumbar and sacral segments of spinal cord were removed to prepare continuous horizontal frozen sections (30 μm) after sedimentation, the sections were dried at room temperature. The corticospinal tract of normal adult rats were shown with Luxol Fast Blue staining method, and observed under Leica DMR microscope. MAIN OUTCOME MEASURES: Positive fibers in Luxol Fast Blue staining. RESULTS: After the Luxol Fast Blue staining, the labeled myelinated nerve fibers were bright blue. They located in the pyramid, decussation of pyramid and the ventral part of posterior funiculus in cervical, thoracic, lumbar and sacral segments of spinal cord. CONCLUSION: Luxol Fast Blue staining method may manifest the distribution of corticospinal tract with clear distinct in adult rats.

Experimental Study on the Effectiveness and Safety of Gardenia Blue Pigment on Anterior Capsule Staining

Objective: To explore the effect of gardenia blue pigment on the anterior capsule staining and its safety performance. Methods: In this study, New Zealand white rabbits were used as the research object to test the effect of gardenia blue pigment on the anterior capsule staining and anterior segment toxicity. In this study, gardenia blue stains of different concentrations (5%, 4%, 3%, 2%, 1%, 0.5%) were prepared to stain the anterior capsule of rabbit crystals, and the effect of the stain was observed to determine the optimal concentration of the stain. Twenty-seven healthy New Zealand white rabbits without eye disease were randomly divided into three groups: gardenia blue staining group;bio-blue positive drug group;physiological saline group. The intraocular pressure, anterior chamber inflammation, corneal endothelial injury, and anterior chamber angle pathological changes were measured in 1D, 7D, and 14D. Results: The results of this study showed that the effect of gardenia blue pigment on the anterior capsule of the lens was the best when the concentration of blue pigment was 2%. There was no change in intraocular pressure in the anterior chamber for a short period of time. There is no damage to the corneal endothelium. Conclusion: The results of this study show that the effect of gardenia blue pigment on the anterior capsule is best when the concentration of blue pigment is 2%, and the stain has no obvious toxic and side effects on the anterior segment of the eye.

A Comparative Study on Four Staining Methods for Antitumor Active Fraction of Periplaneta americana

[ Objective ] This study aimed to compare four staining methods for proteins of SDS-polyacrylamide gel dectrophoresis （SDS-PAGE） and explore a suit- able staining method for the antitumor active fraction of P. americana after polyacrylamide gel electrophoresis. [ Method ] BSA was used as the standard for the comparison of Coomassie brilliant blue staining method, potassium staining method, calcium staining method and silver staining method, on the basis, antitumor ac- tive fraction samples of P. americana were used for SDS-PAGE electrophoresis and staining. [ Result] The results showed that silver staining method could be ac- curately, quickly and easily used for SDS-PAGE staining of the antitumor active fraction of P. amer/cana. [ Conclusion] This study laid the foundation for explo- ring the medicinal value of P. americana.

Can the Prediction of Intrauterine Insemination Results by Used Aniline Blue Stain (ABS) and Sperm Chromatin Dispersion (SCD) Levels?

Introduction: This study aimed to perform routine seminal fluid analysis, sperm DNA fragmentation, and sperm function tests at the chromatin maturation level and evaluate pregnancy in the patients passing intrauterine insemination before starting Intrauterine Insemination (IUI) method. Materials and Methods: In this prospective study, 111 couples who underwent Intrauterine Insemination (IUI) in unexplained infertility patients were admitted to Al-Farah IVF and assisted reproductive center in Baghdad, Iraq between November 2020 and February 2021 were evaluated. Semen fluid analysis was performed based on (WHO 4th) guiding rules. In addition, Sperm Chromatin Dispersion (halo test) and sperm maturation were performed with Aniline Blue Stain (ABS). Results: Sperm Chromatin Dispersion (SCD) groups were compared in terms of pregnancy outcome;the positive pregnancy rate was found to be above in the normal SCD groups (p = 0.0005). In addition, Aniline Blue Stain (ABS) groups were compared in the terms of pregnancy outcome;the positive pregnancy rate was found to be higher in the normal ABS group (p = 0.017). Conclusion: Our study showed that the use of DNA fragmentation (SCD) and sperm maturation tests (ABS) together with routine semen analysis in intrauterine insemination cases will make a significant contribution to the prediction of Intrauterine Insemination (IUI) increased results. So, these results indicate a defect in the effect of DNA fragmentation on the outcome of intrauterine insemination.

大鼠皮质脊髓束Luxol Fast Blue染色

目的:探讨显示大鼠皮质脊髓束的特殊染色方法。方法:选用正常成年SD大鼠延髓和脊髓冰冻切片,应用Luxol Fast Blue染色法进行染色。结果:Luxol Fast Blue染色后,在延髓和脊髓切片中可见灰质呈淡红色,白质呈蓝色,延髓锥体染成深蓝色。锥体中Luxol Fast Blue标记的深蓝色阳性纤维,经锥体交叉后至脊髓灰质后连合背侧,沿脊髓后索腹侧深层下行,至荐段后逐渐消失。在延髓锥体和脊髓颈、胸、腰段后索中,深蓝色的Luxol Fast Blue阳性纤维边界清晰,与周围结构区分明显。结论:运用Luxol Fast Blue染色可清楚显示大鼠皮质脊髓束在脊髓内的定位,是一种简便可靠的皮质脊髓束形态学研究方法。

Evaluation of Fluorescent Stains for Viability Assessment of the Potato Cyst Nematodes <i>Globodera pallida</i>and <i>G. ellingtonae</i>

Potato cyst nematodes (PCNs), Globodera pallida and Globodera rostochiensis, are quarantine pests of potato which cause significant damage to production and farm gate revenue worldwide. Accurately assessing viability of PCN eggs is important for eradication and management programs. The goal of this study was to develop a quick and reliable fluorescent staining method to evaluate viability of G. pallida and Globodera ellingtonae eggs. The staining efficiency of eight fluorescent stains was evaluated using G. pallida eggs compared with the conventional Meldola’s Blue (MB) staining method. The staining efficiency of the fluorescent stains ranged from 80.33 ± 2.99 (Sytox Green) to 100% (Acridine Orange) for non-viable eggs. Two stains were further evaluated for their efficiency in assessing viability of encysted eggs from five different greenhouse-reared G. pallida cyst sources which contained both viable and non-viable eggs. For the G. pallida cyst sources, viability ofencysted eggs were estimated to be 41.02 ± 3.81 to 62.66% ± 3.12% when stained with Acridine Orange (AO) and 79.52% ± 1.54% viability for G. ellingtonae. Both staining time and stain concentration were significant for staining efficiency of released and encysted eggs. Staining time and concentration were optimized for released eggs at 4 h at 10 μg/ml and for encysted eggs at 16 h at 25 μg/ml respectively for AO. Fluorescent stains accurately and rapidly assessed percent egg viability and were determined to be as sensitive as a seven-day incubation with the Meldola’s Blue staining method.

乳酸酚棉兰染色在气传花粉鉴定中的应用

目的探索乳酸酚棉兰(lactophenol cotton blue,LPCB)染色方法在气传花粉镜检中的潜在应用价值。方法分别采用碱性复红(basic fuchsin,BF)染色与LPCB染色鉴定潍坊市春季、夏秋季主要气传花粉并记录每日花粉粒数。结果在鉴定构树花粉、悬铃木属花粉、蒿属花粉、苋科花粉和禾本科花粉方面,LPCB染色较BF染色更具优势,而在鉴定柏科花粉方面,BF染色更具优势。此外,潍坊市夏秋季优势花粉近40年无明显变化,由于绿化树种的改变,悬铃木属花粉已成为本地春季的主要致敏花粉。结论LPCB染色可同时用于真菌和气传花粉的鉴定,在保证鉴定准确性的同时,节约时间成本。

ICG荧光示踪法联合亚甲蓝染色在乳腺癌患者前哨淋巴结活检中的应用价值

目的探讨乳腺癌患者前哨淋巴结活检(SLNB)中应用吲哚菁绿(ICG)荧光示踪法联合亚甲蓝染色的临床价值。方法选择118例乳腺癌患者,以随机数字表法分为2组,对照组、观察组各59例。2组均进行SLNB,术中对照组使用亚甲蓝染色法,观察组使用ICG荧光示踪法联合亚甲蓝染色。比较2组SLN检出情况及血压下降、注射部位感染、坏死、发热等并发症发生情况。结果观察组示踪成功率为98.31%(58/59),高于对照组的86.44%(51/59),差异有统计学意义(P<0.05)。观察组共检出246枚SLN,每位患者平均检出(4.17±0.36)枚;对照组共检出162枚SLN,每位患者平均检出(2.75±0.32)枚;组间对比差异有统计学意义(P<0.05)。2组患者术后均未发生明显并发症。结论与单用亚甲蓝染色相比,联合ICG荧光示踪法在乳腺癌患者SLNB中的应用效果更佳,其示踪成功率更高,检出数量更多,临床应用价值较高。

马尾松木材防霉防蓝变研究进展

马尾松(Pinus massoniana Lamb.)是我国南方分布最广,最主要的用材树种之一,生长速度快、适应能力强、木材力学性能好,被广泛应用于建筑、造纸、装饰、家具、交通等行业。然而,由于其内部含有许多营养物质,易受到霉菌和蓝变菌的侵害。本文重点对国内外传统和新型的马尾松木材防霉防蓝变药剂和处理方法进行了阐述,同时对存在的问题进行了分析,并对研究方向进行了展望,旨在为今后开发新型防霉防蓝变药剂和方法提供理论支持。

基于微创预处理的重组木防霉防腐性能

【目的】研究不同预处理工艺对重组木防霉和防腐性能的影响,旨在寻求最优处理方法以提升重组木在户外多变环境中的使用性能。【方法】采用传统的水煮法、微波法和创新性提出的微创法对重组木进行预处理,利用有机碘化物3-碘代-2丙炔基甲氨酸丁酯(IPBC)对其进行浸渍并采用异氰酸酯胶黏剂(MDI)饰面。通过防霉、防腐试验分析3种处理工艺重组木的防治效力,通过孔隙率、力学性能和尺寸稳定性研究3种处理工艺对重组木质量的影响。【结果】(1)通过微创预处理工艺,重组木的总孔隙率增加了64.1%,载药量提高了88.0%。(2)防霉试验结果显示重组木对照样对黑曲霉和可可球二孢菌的感染等级均为4级,而经过微创预处理/IPBC浸渍工艺的重组木对两种菌的感染等级均为0级,此外通过MDI饰面的重组木抗流失性提高了63.0%。(3)防腐试验结果显示重组木对照样对白腐菌和褐腐菌仅为Ⅱ耐腐等级,而经过微创预处理/IPBC浸渍工艺的重组木对白腐菌和褐腐菌均为Ⅰ耐腐等级。(4)力学和尺寸稳定性试验的结果表明:经过微创预处理/IPBC浸渍工艺的重组木有效保证了优异的力学性能和尺寸稳定性;相比重组木对照样,其抗弯强度和水平剪切强度相比对照样分别提高了14.9%和17.4%,其吸水宽度膨胀率和吸水厚度膨胀率分别降低了25.0%和49.2%。【结论】采用微创预处理/IPBC浸渍工艺制备的重组木对霉菌、变色菌和腐朽菌都具有优异的防治效力,同时提升了力学性能和尺寸稳定性。此外,该微创工艺绿色简单,可有效解决重组木户外应用难题。

小反刍兽疫LAMP反应结果可视化检测方法比较研究

环介导等温扩增(loop-mediated isothermal amplification,LAMP)技术是一种可应用于各种病原体检测、基因分型等领域的等温扩增技术。该技术所需设备简单且易于操作,因此具有较大的发展潜力。根据目前已经存在的判定LAMP结果的可视化方法,以可能对野生小反刍动物造成威胁的小反刍兽疫病毒(Peste des petits ruminants virus,PPRV)cDNA为检测对象,选择合适的引物组,比较评估均可在日光或紫外光下区分阴阳性结果的SYBR Green I、SYBR Safe Stain和羟基萘酚蓝(hydroxy naphthol blue,HNB)3种指示剂方法的适用性与灵敏度。结果表明:3种方法的灵敏度都很高,均与琼脂糖凝胶电泳相当。其中,SYBR Green I结果区分度高、不受体系成分影响且无须额外设备,但成本较高;SYBR Safe Stain具有良好的结果区分度,成本相对较低,但需要额外的紫外照射设备;HNB成本最低,但颜色区分度较低,且易受多种因素影响。建议在选择LAMP指示剂时,综合考虑成本、使用便利性、结果准确性及实验条件,通过充分的体系优化和验证,确保LAMP检测的准确性和可靠性。研究结果突显了不同方法的特征和适用场景,为研究人员根据不同场所与条件选择高效、便捷的LAMP反应结果可视化方法提供参考。

醋酸染色联合蓝激光成像技术对早期胃癌的诊断价值

目的探讨醋酸染色联合蓝激光成像技术对早期胃癌的诊断价值。方法将2021年1月至2023年12月在佛山市顺德区第三人民医院内镜室行电子胃镜检查的300例疑似早期胃癌病例进行研究。所有病例均先后接受白光内镜检查、醋酸染色检查、蓝激光内镜检查、醋酸染色联合蓝激光检查。观察性别、年龄、既往病史、癌症家族史、不良生活习惯、不良饮食习惯等因素对早期胃癌发生的影响,并比较白光内镜、醋酸染色、蓝激光内镜、醋酸染色联合蓝激光内镜4种检查方式对早期胃癌的诊断效果。结果胃癌与非胃癌患者在性别、年龄、胃部疾病史、癌症家族史、吸烟、饮酒、不规律饮食、重盐油腻饮食等因素分布上比较,P<0.05;醋酸染色诊断早期胃癌的灵敏度、准确度均高于白光内镜(P<0.05);蓝激光内镜、醋酸染色联合蓝激光内镜诊断早期胃癌的灵敏度、特异度、准确度均高于白光内镜(P<0.05);蓝激光内镜、醋酸染色联合蓝激光内镜诊断早期胃癌的灵敏度、特异度、准确度均高于醋酸染色(P<0.05);醋酸染色联合蓝激光内镜诊断早期胃癌的灵敏度、特异度、准确度均高于蓝激光内镜,统计学结果有显著差异(P<0.05)。结论诱发早期胃癌的危险因素较多。采取醋酸染色联合蓝激光成像技术诊断早期胃癌效果显著,是一种可行性好的早期胃癌筛查方法。

生物染色剂定位与示踪技术在乳腺外科中的应用

通过对生物染色剂定位与示踪技术在乳腺区段切除术、乳头溢液选择性溢液导管切除术、乳腺钙化灶导丝定位切除术、乳腺癌前哨淋巴结活检术、淋巴结反向示踪引导腋窝淋巴结清扫术/前哨淋巴结活检术、乳腺癌保乳术、腔镜乳腺癌保乳术、腔镜乳腺皮下腺体切除术等术式中的应用进行分类和归纳总结,探讨生物染色剂定位与示踪技术在提高乳腺手术的精准性及微创性方面的优势,旨在为提高乳腺手术的精细化程度及改善术后外观效果提供参考。

紫杉醇和丹皮酚在胶质瘤治疗中的联合抗肿瘤作用

目的探讨分析紫杉醇(Ptx)和丹皮酚(Pae)在胶质瘤治疗中的联合抗肿瘤效果。方法本研究主要采取细胞培养、噻唑蓝(MTT)试验、DAPI染色、细胞流式术等手段,探究通过联合丹皮酚、紫杉醇化疗,能否增强紫杉醇的化疗作用,并减少紫杉醇的化疗剂量,进而减轻其副作用;同时分析Ptx和Pae的协同抗肿瘤作用机制。结果Ptx、Pae两药合用对U87细胞增殖的抑制作用呈剂量依赖性,且不同浓度的Ptx与Pae(125μg/ml)联用时,各浓度组细胞存活率均比单独使用Ptx组显著减少,差异有统计学意义(P<0.05)。表明,Pae协同Ptx促进细胞凋亡可能通过细胞活性氧机制来完成,且Pae协同Ptx可促进细胞ROS的产生及细胞蛋白激酶AKT的磷酸化,并抑制肿瘤的生长。结论Pae能进一步增强Ptx针对胶质瘤细胞的抗肿瘤活性,且Pae联合Ptx主要通过细胞活性氧机制来促进细胞凋亡。

蓝激光成像放大胃镜联合靛胭脂染色对早期胃癌的鉴别价值研究

目的:探讨蓝激光成像放大胃镜(Blue laser imaging magnifying gastroscopy,M-BLI)联合靛胭脂染色鉴别早期胃癌的价值。方法:选取2022年1月至2022年12月医院收治的60例疑似早期胃癌患者作为研究对象,均进行普通白光成像(White light imaging,WLI)检查、M-BLI联合靛胭脂染色检查,以病理检查结果为金标准,分析WLI、M-BLI联合靛胭脂染色对早期胃癌的诊断效能。结果:M-BLI联合靛胭脂染色检查图像质量评分均高于WLI检查(P<0.05)。病理学检查显示,60例疑似早期胃癌患者中,胃癌患者20例,癌前病变患者40例;M-BLI联合靛胭脂染色检查显示,胃癌患者21例,癌前病变患者39例,与病理学检查的Kappa值为0.889;WLI检查显示,胃癌患者20例,癌前病变患者40例,与病理学检查的Kappa值为0.550。M-BLI联合靛胭脂染色诊断早期胃癌的准确度、敏感度分别为95.00%(57/60)、95.00%(19/20)均高于WLI的80.00%(48/60)、70.00%(14/20)(P<0.05)。结论:M-BLI联合靛胭脂染色能获得更清晰的图像,对早期胃癌具有较高的鉴别效能。

几种双向凝胶电泳蛋白质检测方法的比较

本研究对蛋白质染色法进行比较和分析,以求获得满足蛋白质组研究所需的适宜的蛋白质检测法。实验采用2-D电泳分离急性早幼粒白血病细胞株NB4的全蛋白及1-D电泳分离蛋白质分子量标准物,从蛋白质检测的灵敏性、质谱兼容性、操作的简便性等角度,比较了传统考马斯亮蓝染色法、胶体考马斯亮蓝染色法、改良考马斯亮蓝染色法和银染法4种蛋白质着色法对凝胶分离蛋白质的检测影响,探讨了影响蛋白质染色与鉴定的影响因素。结果显示,银染检测灵敏度最高,但质谱鉴定相容性差,鉴定率近10%;改良考马斯亮蓝染色法检测灵敏度也高,可达4ng/spot,质谱鉴定率可接近55%。结论:改良考马斯亮蓝染色法的蛋白质检测灵敏度高,与质谱兼容好,能满足蛋白质组研究需要。

一种简便的考马斯亮蓝G250蛋白质染色方法

介绍一种快速、简便、几乎无背景的考马斯亮蓝G2 5 0 (CBBG2 5 0 )染色方法 .该方法所用试剂仅为稀盐酸和CBBG2 5 0 ,CBBG2 5 0的工作浓度为 0 0 0 15 % ,灵敏度达 0 0 2 μg/带 ,染色 2h达 70 % ,4h以上或染色过夜即可充分染色 .与以往的考马斯亮蓝染色方法相比 ,该方法有经济方便、灵敏度高、几乎无背景等优点 。

木材蓝变色生物控制研究的现状

木材蓝变是边材变色最常见的一种真菌性变色,主要由子囊菌纲的Ceratocystis、Ophiostoma、Ceratocystiopsis属和半知菌纲的Aureobasidium、Rhinocladiella属真菌危害引起的,多发生在马尾松、杉木、橡胶木及西南桦等树种的木材上。木材蓝变严重影响了木材的美观。采用化学方法防治木材蓝变,虽有一定效果,但造价高也不环保。应用生物防治木材蓝变,是通过直接抑制蓝变菌的生长来达到防止蓝变的发生。生物防治的技术关键是控制菌种的筛选、代谢产物的生物控制、菌丝寄生的控制、以及环境条件对控制作用的影响等。