简易革兰氏染色法及其质量控制

为了使染色方法更简便、快速，我们采用了革兰染色３步法，效果和四步法一样，现将革兰染色３步法与质量控制报告如下。１材料和方法１．１材料１．１．１结晶紫染色液：（１）甲液：结晶紫２ｇ，９５％乙醇２０ｍｌ。（２）乙液：草酸铵０．８ｇ，蒸馏水８０ｍｌ。甲乙２...     

革兰氏染色三步法

革兰氏染色法仍是目前临床细菌学检验最常用的检测方法 ,该法为鉴别染色法 ,经染色可将革兰氏阳性菌染成紫色 ,革兰氏阴性菌染成红色。二者对比鲜明 ,结果准确可靠。革兰氏染色法通常为四步法 ,经过长期使用 ,认为此法比较复杂 ,把四步法改为三步法 ,具有省时省事等特点 ,因此 ,对这一方法的适用性作了进一步的实验观察。1　材料与方法1 1　材料1 1 1　结晶紫染色液　甲液 :结晶紫 2ｇ ,95 %乙醇 2 0ｍｌ;乙液 :草酸铵 0 .8ｇ ,蒸馏水 80ｍｌ。甲乙二液先分别溶解 ,然后混合在一起 ,过滤除去残渣后装入滴瓶中备用。1 1 2　碘液　碘 1ｇ ,碘…     

介绍一种革兰氏染色液的改良配方

<正> 1.染色液配方1·1 结晶紫液 结晶紫0.5g加95％乙醇50ml溶解,再加入1g/dL草酸铵溶液450ml混匀。1·2 革兰氏碘液 20g碘化钾溶于30ml蒸馏水中,加碘片10g,全部溶解后加蒸馏水至300ml混匀。1·3 沙黄溶液 0.5g沙黄加95％乙醇35ml,溶解后加蒸馏水至 500ml,混匀。     

肿瘤坏死因子细胞毒活性检测方法比较

比较MTT比色分析法、中性红摄取法、结晶紫染色湿测法、结晶紫染色干测法和~3H-TdR摄入法五种方法检测L929细胞毒活性的敏感性和稳定性。结果表明五种方法检测的TNF活性有明显差异,以结晶紫染色干测法具有较敏感、稳定、简便之优点.并用此方法检测TNF对三种不同肿瘤细胞的细胞毒活性结果,不同的肿瘤细胞对TNF的敏感性有明显差异,提示可用结晶紫染色干测法检测TNF对不同肿瘤细胞的细胞毒性效应的敏感性。以供临床治疗参考。     

用高铁氰化血红蛋白标准液间接进行电子血球计数器标准定值(摘要)

市售氰化血红蛋白标准液,不能直接用于电子血球计数器的标准定值,作者用高铁氰化血红蛋白液在721型分光光度计上制备标准曲线后,任选一血液标准液同时进行比色法和电子血球计算器血红蛋白测定(分别平行测定五份,取平均值),将测定值定在电子血球计算器上。工作中可随时用本法对电     

一种实用的厌氧菌芽胞形态结构片制作法

目的 探讨一种实用的厌氧菌芽胞形态结构片制作法.方法 以改进法分离培养产芽胞梭菌,采用革兰染色液前3种,即结晶紫液,卢戈碘液,95%酒精染色.结果 细菌菌体及芽胞壁被染成紫色,芽胞为清晰透明的卵圆形和圆形空泡,大于菌体,位于次末端,片子保质期长.结论 该方法简单,效果良好.     

细胞学涂片快速染色法

细胞学涂片一般采用常规 HE 染色。笔者介绍了一种快速染色方法,此方法可用于宫颈涂片、痰涂片、穿刺物涂片、新鲜组织印片等细胞学检查。1 材料和方法1.1 试剂配制 (1)2%结晶紫染液:结晶紫2g,蒸馏水100ml。先将结晶紫加入已加热     

SARS-CoV细胞空斑试验

目的 建立适用于生物安全三级实验室操作的SARS-CoV空斑试验方法.方法 SARS-CoV10倍系列稀释接种Vero-E6细胞,用0.6%琼脂糖覆盖中性红着色(琼脂糖-中性红法)或覆盖1%甲基纤维素,4%甲醛固定,结晶紫染色(甲基纤维素-结晶紫法),空斑计数.结果 琼脂糖-中性红法病毒滴度为6.14 Lg pfu/mL,甲基纤维素-结晶紫法病毒滴度为6.57 Lg pfu/mL.结论 二种空斑试验方法相比,病毒滴度无明显差异.甲基纤维素-结晶紫空斑试验法形成的空斑比琼脂糖-中性红法清晰、操作简单安全、结果可长期保存.     

自动血球计数器的国产LY-1洗净液和稀释液

为解决PC-603血球计数器与相应的方法学试剂配套问题,北京生化仪器厂根据PC-603血球计数器的红白血球、血红蛋白计数要求及小孔管孔径的特殊要求,较成功地摸索出合理的洗净液和稀释液配方及配制方法,制出了LY-1洗净液和稀释液。经实验室分析和临床试用证实,各项指标基本与日本原装溶液相似,可供临床广泛使用。 LY-1洗净液无气味,粘稠度小,能溶     

血小板染色后计数方法

在普通光镜了计数血小板，由于对比度较差，容易造成视力疲劳。我们采用血小板预染色后计数。用3种不同染色液显示血小板呈不同青绿色，结果皆不影响血小板计数，报道于下。1材料与方法1．1配制方法：取草酸铵1g，Na2EDTA0．1g，甲醛0．1ml，加水100ml，加亚甲蓝1g，煌焦油蓝1g     

Shi Da-zhuo (史大卓) —An outstanding cardiologist of integrative medicine

Prof. SHI Da-zhuo, Ph.D., male, was born on March 20, 1960. Prof. SHI entered the Ph.D. program in 1990 at the China Academy of Chinese Medical Sciences under the supervision of Prof. CHEN Ke-ji, majoring in the treatment of cardiovascular diseases. After receiving his Ph.D. degree in 1993, Prof. SHI started working at the Cardiovascular Center in Xiyuan Hospital affiliated to China Academy of Chinese Medical sciences.     

Dr.Zhang Ren's Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

Cloning and sequence analysis of β-actin gene from Aedes albopictus （Diptera： Culicidae）

Objective: To obtain the complete β-actin gene from Aedes albopictus. Methods: Total RNA was extracted from C6/36 cells. Degenerate primers were designed based on the β-actin sequences of An. gambiae, Ae. aegypti, Cx. pipiens pallens and D.melanogaster. By RT-PCR, the product was amplified, purified, cloned into the pGT vector and sequenced. The β-actin sequence was aligned and phylogenetically analyzed by the BLAST program and the CLUSTAL W program. Results: A sequence of 1132 bp including an open reading frame of 1131 bp was obtained (GenBank DQ657949). The deduced protein had 376 amino acids.Aligned to SWISS-PROT, it exhibited a high level of identity with β-actins from Anopheles, Drosophila and Culex at the amino acid sequence level. Phylogenetic analysis indicated that Ae. albopictus β-actin was much more homologous with invertebrate β-actin than with vertebrate β-actin. Conclusion: The gene may be used as the internal control in the experiments of Ae. albopictus.     

Role of myeloid differentiation factor 88 in HSP60 signal transduction in dendritic cells

Objective: To explore the role and mechanism of myeloid differentiation factor88 (MyD88) in HSP60 signal transduction in dendritic cells. Methods:Mouse DCs were cultured from murine bone marrow cells. The DC marker CD11c was detected by flow cytometry, then DCs were divided into control group, HSP60 groupand RNA interference group. Control group was cultured under normal condition, and HSP60 group was cultured with 10 μg/ml of HSP60. RNA interference group was first cultured with MyD88 siRNA forl2 hours and then HSP60 was added into the culture mixture. All groups were cultured for 48 hours. Immunochemistry was used to detect the concentration of MyD88 and NF- κB. Western blot was used to detect the concentration of MyD88. Flow cytometry and mixed lymphocyte reaction (MLR) were used to detect the phenotype and functional properties of DCs. ELISA was used to detect the concentration of TNF-α, IFN-γ and IL-12 in the supernatant. Results:The expression of CD11c in marine bone marrow DCs was 88.76%. HSP60 stimulation increased the expression of CD80, CD86, MHC-Ⅱ in DCs and TNF-α, IFN-γ, IL-12 secretion in the supematant. HSP60 stimulation also increased the level of MyD88 in the cytoplasm and promoted the shift of NF-κB to karyon and the proliferation of allogeneic T cells. MyD88 siRNA could decreaseMyD88 and inhibit these effects induced by HSP60. Conclusion:HSP60 activates DCs through MyD88-dependent pathway. MyD88 plays a critical role in HSP60 signal transduction. Inhibition of MyD88 may be a novel way for treating disease correlated with HSP60.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

股骨远端粉碎性骨折双板固定的生物力学研究

目的探讨治疗股骨远端粉碎性骨折的新方法。方法应用10件成人鲜尸股骨,造成股骨远端髁间、髁上粉碎性骨折,内侧采用聚乙烯板、外侧采用钢板双向固定,分别作压缩、弯曲、扭转等生物力学实验研究。另取10件标本进行单向外侧钢板固定作为对照。结果双向固定组抗弯曲、扭转能力显著增强(P<0.01),抗压缩能力也较单侧固定组强(P<0.05)。结论双向固定治疗股骨远端粉碎性骨折有明显的应用价值。     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

REASSESSMENT OF DEFIBRASE IN TREATMENT OF ACUTE CEREBRAL INFARCTION： A MULTICENTER, RANDOMIZED, DOUBLE-BLIND,PLACEBO-CONTROLLED TRIAL

Objoctive To evaluate the efficacy and safety of defibrase in patients with acute cerebral infarction by a large sample, multicenter, randomized, double-blind, placebo-controlled clinical trial. Mothods Patients with acute cerebral infarction within 12 hours of stroke onset were randomly assigned to receive either an initial intravenous infusion of defibrase 15 U plus normal saline 250 mL or 250 mL of normal saline only. Subsequent infusions of defibrase 5 U or placebo （normal saline） were given on the 3rd, 5th, 7th, and 9th day, respectively. Both groups received standard care of acute cerebral infarction. The primary efficacy outcome was functional status （Barthel Index） at 3 months after treatment. Safety outcome were bleeding events and mortality rate. Secondary outcome included Chinese Stroke Scale （CSS） score at 14 days and recurrence rate of stroke at 1 year. A total of 1053 patients were enrolled at 46 centers from September 2001 to July 2003, and 527 patients were randomly assigned to receive defibrase and 526 to receive placebo. A similar proportion of patients in both groups completed a full course of treatment. There was a significantly greater proportion of favorable functional status （Barthel Index 1≥95） in defibrase group than in placebo group at 3 months （52.2% vs. 42.8%, P 〈 0.01）, and the proportion of dependent functional status （Barthel Index ≤60） was a little lower in defibrase group compared with placebo group （27.7% vs. 32.4%）. These differences were more obvious among patients who were treated within 6 hours of stroke onset. Patients in defibrase group had better improvement with respect to CSS score than those in placebo group at 14 days （P 〈 0.05）. Recurrence rate of stroke at 1 year was lower in the defibrase group compared with placebo group （6.2% vs. 10.1%, P = 0.053）. Patients in defibrase group had higher risk of extracranial bleeding events （4.7% vs. 1.5%, P 〈 0.01 ） and a tendency of higher risk of symptomatic intracranial hemorrhage. The hemorrhage incidence was higher in patients with fibrinogen level 〈 130 mg/dL than ≥ 130 mg/dL （10.6% vs. 3.8%, P 〈 0.05）. Mortality rate at 3 months were slightly higher in defibrase group than placebo group （5.9% vs. 4.2%）. Conclusions The defibrase is effective to improve neurological function and function of daily living for patients with acute cerebral infarction within 12 hours of symptom onset. The efficacy was even better for acute cerebral infarction within 6 hours of onset. The increased risks of intra- and extracranial hemorrhage during defibrase administration were related to the plasma fibrinogen level.