新型隐球菌脑膜炎53例临床分析

目的总结分析新型隐球菌性脑膜炎的临床特征。方法分析53例新型隐球菌脑膜炎患者的临床表现、实验室检查、治疗及转归。结果新型隐球菌脑膜炎的临床表现以颅内压增高、脑膜刺激征为主。有基础性疾病者30例(占56．6%)。发病前有鸽子、禽类接触史者30例。脑脊液压力升高51例,其中49例超过200 mmH_2O。30例患者应用两性霉素B与氟康唑联合治疗。53例患者治愈4例,好转27例,死亡5例,自动出院17例。结论新型隐球菌性脑膜炎的主要临床表现是进行性颅内压增高。对进行性颅内压增高、脑膜刺激征阳性的患者反复进行脑脊液墨汁染色查新型隐球菌,有助于减少该病的误诊和误治。两性霉素B联合氟康唑静滴治疗隐球菌脑膜炎仍然安全、有效。     

获得性免疫缺陷综合征合并新型隐球菌脑膜炎临床分析

目的总结分析新型隐球菌性脑膜炎的临床特点,提高对获得性免疫缺陷综合征(AIDS)合并隐球菌性脑膜炎的认识。方法分析总结住院的72例患者的临床特点,在给予高效抗反转录病毒治疗的同时,给予抗真菌治疗。结果 AIDS合并隐球菌性脑膜炎的临床表现以颅内压增高(81.9%),脑膜刺激征为主(84.7%),全部合并有概率性感染(100%),免疫功能低表现为CD4+<50 cell/μL(83.3%)。结论 AIDS合并隐球菌性脑膜炎病情重、预后差、病死率高(37.5%),反复实验室病原学检查结果是诊断新型隐球菌性脑膜炎的主要依据。两性霉素B联合氟胞嘧啶治疗新型隐球菌性脑膜炎仍是安全、有效的。     

新型隐球菌性脑膜炎15例误诊原因分析

近年来 ,随着抗生素、免疫抑制剂和皮质激素的广泛应用 ,机体菌群失调或免疫功能下降 ,致使新型隐球菌性脑膜炎(隐脑 )的发病率明显升高。 1985～ 2 0 0 2年 ,我院误诊 15例新型隐球菌性脑膜炎 ,回顾分析如下。1　临床资料1.1　一般资料　 15例隐脑中 ,男 9例 ,女 6例 ;年龄 3～ 12岁。发病前有慢性疾病史者 2例 ,1例为白血病 ,1例为肾病综合征。1.2　误诊情况　①误诊为结核性脑膜炎 3例。患者起病均缓慢 ,低热、头痛、恶心、呕吐 ,脑膜刺激征阳性 ;腰穿脑脊液压力高 ,细胞数 10 0～ 30 0× 10 6/L ,蛋白升高 ,糖和氯化物降低。因患者临床…     

隐球菌脑膜炎42例临床分析

目的：归纳分析42例隐球菌脑膜炎的发病情况、临床特点并总结抗真菌药物的治疗经验,以提高对隐球菌脑膜炎的诊治水平。方法：回顾分析四川大学华西医院2001～2007年所收治42例隐球菌脑膜炎病例的临床表现、实验室检查结果、抗真菌药物的疗效及预后,并对两性霉素B联合5-氟胞嘧啶抗真菌治疗的药物剂量、疗效、不良反应以及疗程与预后的关系加以剖析。结果：临床以发热、头痛、颅内压升高、脑膜刺激征为主要表现,采用两性霉素B联合5-氟胞嘧啶治疗,总有效率78.5%,42例患者治愈9例,好转24例,死亡5例,自动出院后失访4例。结论：隐球菌脑膜炎由于临床表现、脑脊液常规和生化检查以及影像学检查无明显特异性,易于误诊;脑脊液墨汁染色有助于早期诊断本病,对疑似病例反复进行脑脊液墨汁染色有利于确诊;两性霉素B联合5-氟胞嘧啶治疗治疗隐球菌脑膜炎效果良好。但需注重合理应用并密切监测不良反应。     

新型隐球菌性脑膜炎12例临床分析

目的：探讨新型隐球菌性脑膜炎（CM）的临床特点，诊断及治疗方法。方法：对12例CM患者的临床资料进行回顾性分析。结果：12例患者中，8例为急性及亚急性起病，4例起病匿隐，临床上主要表现为头痛，颅高压明显，脑膜刺激症阳性，脑脊液改变与结核性脑膜炎相似；CM误诊率高；本组患者经联合治疗疗效明显。结论：CM是以头痛、颅高压和脑脑膜刺激症为特点，误诊率高，反复腰穿检查有助于早期诊断。     

46例新型隐球菌性脑膜炎临床诊治分析

目的分析新型隐球菌脑膜炎的易感因素、临床特点、治疗及转归。方法回顾性分析2000年12月至2011年10月确诊的46例新型隐球菌脑膜炎。总结其临床表现、实验室检查、治疗及转归情况。结果46例患者均经脑脊液墨汁染色阳性确诊。22例（47．83％）合并有其他免疫系统疾病。治愈9例、好转18例、6例死亡、13例自动出院。结论患有免疫系统疾病是新型隐球菌脑膜炎的易感因素之一,在患有免疫系统疾病患者中出现发热、头痛等脑膜刺激征阳性,应及时行腰穿,反复行脑脊液墨汁涂片找隐球菌,及时明确诊断,联合鞘内注射两性霉素B能缩短脑脊液隐球菌转阴时间,以提高隐球菌脑膜炎临床疗效。     

新型隐球菌脑炎40例死因分析

目的研究新型隐球菌脑膜炎（CM）死亡原因并讨论降低死亡的对策。方法对我院1965—2005年确诊为隐球菌脑膜炎84例中住院期间死亡40例（47．6％）的临床资料进行回顾性分析。结果（1）主要症状体征依次为头痛、呕吐、脑膜刺激征、发热、瞳孔不等大、视乳头水肿以及意识障碍。（2）脑脊液改变：颅内压增高39例中〉3．92kPa24例（61．5％）；墨汁涂片，32／34例阳性（94．1％）：隐球菌培养16／19例阳性（84．2％）。（3）用两性霉素B或大蒜液治疗。少数病例加用5-氟胞嘧啶或氟康唑。结论抓住主要症状体征和脑脊液检查，及时明确诊断。尽快使用足量两性霉素B和5-氟胞嘧啶或氟康唑联合治疗，多途径给药。可减少不良反应与并发症，降低死亡率。     

二性霉素B治疗小儿新型隐球菌脑膜炎的护理

新型隐球菌脑膜炎是新型隐球菌侵犯中枢神经系统引起的脑膜炎，该病原体通过呼吸道及皮肤破损处进入血液循环感染中枢神经系统。起病隐匿，临床表现以头痛为首发症状，伴有发热、脑膜刺激征，甚至抽搐、昏迷本病如不及时治疗常于数日内死亡。我科采取了二性霉素B联合其他药物治疗小儿新型隐球菌脑膜炎，取得了较好的疗效，现将护理体会总结如下。     

新型隐球菌脑炎/脑膜炎4例临床分析并文献复习

目的:报告新型隐球菌脑炎/脑膜炎4例,并结合文献检索探讨其临床特点及可能的发病机制。方法:回顾分析4例病原学诊断明确的新型隐球菌脑炎/脑膜炎的临床资料,结合文献检索探讨其发病特点及可能发病机制。结果:患者主要表现为头痛、发热、恶心、呕吐及意识障碍;4例均在脑脊液培养中发现新型隐球菌,其中2例墨汁染色发现新型隐球菌;免疫力低下患者易感;脑脊液压力高;主要与结核性脑炎/脑膜炎鉴别;常用的临床治疗方案是两性霉素B联合氟胞嘧啶诱导治疗,氟康唑维持及巩固治疗。结论:中枢神经系统隐球菌病临床表现无特异性,脑脊液墨汁染色阳性率低,在临床诊断过程中对疑似患者,需要反复多次行脑脊液墨汁涂片及培养,减少患者误诊和漏诊。     

1例新型隐球菌型脑炎病人的护理

隐球菌脑膜炎主要是由新型隐球菌通过各种渠道侵入颅内引起的脑膜炎或脑膜脑炎。是最常见的颅内真菌感染。主要的临床症状为头痛、发热、头晕、呕吐、视力模糊或复视等。脑脊液检查做墨汁染色可在显微镜F看到其菌体，病理学检查可见脑膜刺激症状及眼底的视乳头水肿，腰椎穿刺结果提示颅内高压。     

改进酶消化法培养SD大鼠成骨细胞及其鉴定

摘要 目的：改进SD大鼠成骨细胞的体外分离、培养方法并进行功能鉴定。方法：将新生SD大鼠处死，无菌条件下取出颅骨，剔净骨膜后剪成1mm3大小组织块。组织块经0.25%胰蛋白酶消化20min，继以0.1% II型胶原酶消化60 min，收集上清离心，所得成骨细胞接种于培养瓶中并行“多次贴壁法”纯化。观察细胞形态学，选用ALP Gomori 钙钴法染色，钙结节茜素红法染色及I型胶原免疫组化染色等方法进行鉴定。 结果：培养的细胞具有典型的成骨细胞形态特征，在体外培养时可维持其在体内的功能:合成碱性磷酸酶、形成矿化结节，I型胶原染色阳性。结论：用改进后的酶消化法分离、培养SD大鼠成骨细胞，更能减少胰蛋白酶在消化过程中对细胞造成的损伤，所获成骨细胞量多、纯度高，操作简易可行，可作为骨组织工程种子细胞常规的培养方法。     

雷尼替丁三联疗法治疗消化性溃疡疗效观察

目的：雷尼替丁三联疗法与奥美拉唑三联疗法治疗消化性溃疡的疗效比较。方法：将73例消化性溃疡随机分为两组。治疗组：37例,雷尼替丁150 mg、阿莫西林1000 mg、甲硝唑400 mg每日2次,治疗2周后,单用雷尼替丁150 mg连用4周。对照组：36例,奥美拉唑20 mg、阿莫西林1000 mg、甲硝唑400 mg每日2次口服。治疗2周后,单用奥美拉唑20 mg连用4周。治疗期间每周到门诊随访,记录临床症状改善情况。用药结束后1月做胃镜检查。结果：治疗后两组的临床症状改善或消失。胃镜复查结果无统计学差异。结论：治疗组和对照组的疗效相同。     

A gut-specific serine protease from the malaria vector Aanopheles gambiae is downregulated after blood ingestion

A chymotrypsin-like serine protease gene (AgChyL) was cloned from the mosquito Anopheles gambiae by a polymerase chain reaction (PCR)-based subtractive cDNA cloning strategy. AgChyL messenger RNA (mRNA) is abundant in the adult female gut prior to, and for 8 h following, a blood meal. During the peak of digestion, from 12 to 24 h following a blood meal, AgChyL mRNA abundance decreased to barely detectable levels. AgChyL mRNA was abundant again by 48 h following a blood meal. Recombinant pro-AgChyL was expressed in Escherichia coli. The pro-enzyme can be activated by trypsin. Activated AgChyL cleaves the synthetic chymotrypsin substrate succinyl-L-Ala-Ala-Pro-Phe-nitroanilide, but not two other synthetic chymotrypsin substrates or synthetic trypsin and elastase substrates. The potential role of AgChyL in the coordination of An. gambiae digestion is discussed.     

Rapid acid digestion and simple microplate method for milk iodine determination

Iodine deficiency leads to deficiency of thyroid hormones, which causes mental retardation in infant. Laboratory confirmation is important in its diagnosis. The major problems associated with the existing methods for iodine determination in milk samples are: 1) nonsafe alkaline solution; 2) harsh thermal condition; and 3) extra time required to complete various steps. In this study, a simple and rapid colorimetric method was investigated, which used acid digestion in combination with a rapid microplate reading format method to determine the total iodine content in milk. Sample digestion was done on 50 microL milk in metavanadate/perchloric, at 230 degrees C for 10 min. After digestion, iodine determination was based on the Sandell-Kolthoff reaction. The reaction results were read in 96-well microplates by an enzyme-linked immunosorbent assay (ELISA) reader. The determination range of the assay was between 2 and 40 microg/dL. The within-run coefficient of variation percent in three levels (3, 12, and 36 microg/dL) ranged from 6.7 to 9.3 and between-run coefficients of variation ranged from 8.6 to 12.3%. The results obtained (n=70) by the optimized method have good correlation with the results of alkaline incineration as a reference method (n=70; r2=0.907; y=0.952x+1.77). Recovery tests for accuracy assessment in six levels from 6.2 to 34.2 microg/dL) were between 91.3 and 113%. This method has enabled us to achieve 0.12 microg/dL sensitivity. The results of this study show that a quick acid digestion combined with mild thermal and low sample volume with a quick reading of assay results were the main advantages of the acid digestion and microplate reading format.     

Effect of in vitro non-enzymatic glycosylation of human skin collagen on susceptibility to collagenase digestion

The effect of glycosylation on susceptibility of skin collagen to collagenase digestion was studied in a skin sample obtained at autopsy from the interscapular region of a 24 year old white male who had died of an acute illness and who had no history of diabetes. Homogeneous suspensions of insoluble collagen were prepared, and were incubated in 50 mmol l-1 dextrose at pH 7.35 and 37 degrees C for 7 days. Non-enzymatic glycosylation measured by the weak acid hydrolysis/thiobarbituric acid method increased from 13.1 +/- 1.0 (n = 5) to 45.2 +/- 5.5 (n = 8) nmol fructose per 10 mg collagen (P less than 0.001). Digestion of collagen using clostridial collagenase was monitored by measuring (a) hydroxyproline content and (b) absorption at 206 nm of the supernatant after centrifugation to remove substrate. The rate of digestion was similar in glycosylated and control collagen. We conclude that the ketoamine link formed in non-enzymatic glycosylation does not increase the resistance of collagen to enzymatic digestion. The possibility remains that subsequent rearrangement of this link could be important in this respect.     

Cloning and characterization of a gut-specific cathepsin L from the aphid Aphis gossypii

We have characterized proteinase activities in gut extracts from the cotton-melon aphid (Aphis gossypii Glover), an insect feeding strictly on protein-poor phloem. The major, if not exclusive, intestinal proteinases of this aphid are of the cysteine type. A cDNA has been cloned from a gut library and codes for the cysteine proteinase AgCatL, a cathepsin L-like cysteine proteinase. The AgCatL protein shows high sequence similarity with mammalian and some arthropod cathepsin L-like proteinases, but can be reliably distinguished from the secreted (digestive) proteinases identified in other arthropods. AgCatL is widely expressed in aphid intestinal cells. Immunolocalization of AgCatL showed an intense signal at the level of the anterior 'stomach' part of the midgut, and especially at intracellular localization. Although the precise role of AgCatL in aphid midgut physiology is still unclear, this enzyme could be involved in the processing of exogenous ingested polypeptides.     

Criteria for consistent and high sensitivity of dna in situ hybridization on paraffin sections: Optimal proteolytic enzyme digestion

It is technically challenging for the detection of target DNA in low abundancy, such as viral DNA sequences in latently infected cells by nonisotopic in situ hybridization (ISH). Consistent result is even more difficult to achieve on routine paraffin sections. Proteolytic enzyme digestion is most critical for both consistency and sensitivity of the technique. We here have investigated the effect of enzyme digestion on cell morphology, protein and DNA reduction, and hybridization efficiency. The results demonstrated that enzyme digestion improves efficiency of ISH through a process involving partial DNA purification on sections. There is a clear relationship between proteolytic enzyme digestion, morphology changes, and hybridization efficiency. Although detection of DNA sequences in abundance can be achieved within a relatively wide range of digestion levels, maximum hybridization efficiency was always related to the cells, which showed morphology of nuclear swollen, weak homogeneous chromatin staining with hematoxylin and loss of visible nuclear membrane. Detection of viral DNA in low copy number critically depends on the creation of the morphologic changes by enzyme digestion. The morphological changes would therefore serve as important criteria for optimal digestion, result interpretation, and comparison.     

Gastrointestinal digestion of food allergens: effect on their allergenicity.

This paper reviews the in vitro digestion models developed to assess the stability digestion of food allergens, as well as the factors derived from the methodology and food structure that may affect the assay results. The adequacy of using the digestion stability of food allergens as a criterion for assessing potential allergenicity is also discussed. Data based on the traditional pepsin digestibility test in simulated gastric fluid are discussed in detail, with special attention to the influence of the pH and pepsin: allergen ratio in the pepsinolysis rate. This review points out the importance of using physiologically relevant in vitro digestion systems for evaluating digestibility of allergens. This would imply the sequential use of digestive enzymes in physiological concentrations, simulation of the stomach/small intestine environment (multi-phase models) with addition of surfactants such as phospholipids or bile salts, as well as the consideration of the gastrointestinal transit and the effect of the food matrices on the allergen digestion and subsequent absorption through the intestinal mucosa. In vitro gastrointestinal digestion protocols should be preferably combined with immunological assays in order to elucidate the role of large digestion-resistant fragments and the influence of the food matrix on the stimulation of the immune system.