青蒿素衍生物抗卡氏肺孢子虫体外作用的研究

目的 研究不同浓度青蒿素衍生物双氢青蒿素、青蒿琥酯对体外培养卡氏肺泡子虫的作用。方法 接种卡氏肺孢子虫（Pc）于人肝癌细胞系（HepG-2)。4h加入试验药物和对照试剂。双氢青蒿素药物浓度分别为：100、50、10、5、0．5μmol/L；青蒿琥酯药物浓度分别为：100、50、10、5μmol/L；对照药喷他脒：15μmol/L。以后每隔1d取培养液观察，分别作六亚甲基四胺银（GMS）和Diff-Quik(DQ)染色计数Pc包囊和滋养体数目。结果 Pc滋养体和包囊在体外培养细胞上的生长高峰出现于第5d，双氢青蒿素浓度为50μmol／L和100μmol／L时对Pc滋养 的抑制率分别为88．0％与90．1％，与喷他脒15μmol/L的抑制率90．0％相当；而青蒿琥酯浓度为100μmol/L时方可达到Pc滋养体抑制率83．4％。Pc包囊抑制率，双氢青蒿素和青蒿琥酯浓度为100μmol/L分别为67．5％和67．3％，与喷他脒的抑制率75．0％之间无统计学差异。结论 双氢青蒿素和青蒿琥酯在一定浓度时67．3％，对Pc的抑制作用与喷他脒相当；而双氢青蒿素的抑制作用略高于青蒿琥酯。两者对Pc滋养体的抑制率均高于对Pc包囊。     

瑞香素体外抗卡氏肺孢子虫作用的初步研究

目的研究瑞香素在体外抗卡氏肺孢子虫(Pneumocystiscarinii,Pc)的作用。方法建立Wistar大鼠卡氏肺孢子虫肺炎模型;分离、纯化虫体,进行体外培养。测定不同浓度的瑞香素体外抗Pc的作用,观察经FeSO4、MgSO4螯合后的瑞香素对虫体的生长影响情况。用透射电镜观察药物处理后的虫体超微结构改变。结果瑞香素抗Pc作用效应在1～20μmol/L浓度范围呈剂量依赖和时间依赖关系。10μmol/L瑞香素与1.0μg/ml喷他脒对Pc生长影响效果相当。若预先将瑞香素与FeSO4按21比例混合后,瑞香素对Pc的生长抑制作用大大减弱。瑞香素处理后的虫体的超微结构有改变。结论瑞香素在体外对Pc生长有抑制作用。     

白果内酯治疗实验大鼠肺孢子虫肺炎电镜观察

目的　观察白果内酯对卡氏肺孢子虫超微结构的影响。方法　地塞米松连续皮下注射Wistar大鼠 6周 ,建立大鼠卡氏肺孢子虫肺炎动物模型 ,腹腔内注射白果内酯 30mg/(kg·d)× 8d。停药 1周后取大鼠肺组织作超薄切片 ,透射电镜观察。结果　白果内酯作用后的卡氏肺孢子虫虫体有大量空泡形成 ,细胞器肿胀破坏 ,髓样结构形成 ,胞膜破坏 ,胞质内出现电子密度较高颗粒。结论　白果内酯可破坏卡氏肺孢子虫的超微结构 ,从而引起虫体死亡。     

卡氏肺孢子虫包囊抗原纯化方法的研究

目的　研究卡氏肺孢子虫包囊抗原的纯化方法。　方法　肌肉注射地塞米松诱发大鼠卡氏肺孢子虫肺炎 ,瑞 -姬氏染液复合染色法检获肺组织中的肺孢子虫包囊 ,用改进的胶原酶消化法、Percoll不连续密度梯度离心分离、超声粉碎纯化卡氏肺孢子虫包囊抗原。　结果　采用浓度为 0 .0 5 %胶原酶消化 2次 ,30 min/次 ,在 1.0 30 g/ ml密度层可获得相对纯净而完整的卡氏肺孢子虫包囊抗原。　结论　本研究可提供高纯度卡氏肺孢子虫抗原 ,为进一步建立对卡氏肺孢子虫肺炎的有效诊断方法具有十分重要的意义。     

卡氏肺孢子虫包囊抗原纯化方法的研究

目的 研究卡氏肺孢子虫包囊抗原的纯化方法。方法 肌肉注射地塞米松诱发大鼠卡氏肺孢子虫肺炎，瑞姬氏染液复合染色法获肺组织中的肺孢子虫包囊，用改进的胶原酶消化法，Percoll不连续密度梯度离心分离，超声粉碎纯化卡氏肺孢子虫包囊抗原。结果 采用浓度为0．05％胶原酶消化2次，30min/次，在1.030g/ml密度层可获得相对纯净与而而完整的卡氏肺孢子虫包囊抗原。结论 本研究可提价蒿纯度卡氏肺孢子虫抗原，为进一步建立对卡氏肺孢子虫肺炎的有效诊断方法具有十分重要的意义。     

经双氢青蒿素治疗后卡氏肺孢子虫肺炎大鼠的肺部病理学变化

目的　研究经双氢青蒿素治疗后卡氏肺孢子虫肺炎 (PCP)大鼠肺部病理学变化。方法　以地塞米松磷酸钠皮下注射Wistar大鼠建立卡氏肺孢子虫肺炎动物模型 ,用 6 0mg/kg双氢青蒿素治疗实验大鼠 ,杀鼠取肺 ,用光镜和电镜观察肺部病理学变化 ,同时设有感染对照组和正常对照组。结果　肺印片中卡氏肺孢子虫 (Pc)包囊数目显著减少 ,肺组织炎症明显减轻 ,Pc滋养体表膜和核膜破裂 ,胞质中出现大量空泡和高电子密度颗粒 ,Pc包囊中也出现空泡 ,囊内小体变性坏死。结论　双氢青蒿素可杀死Pc滋养体和包囊 ,从而减轻肺组织的炎症反应。     

青蒿素衍生物体外抗卡氏肺孢子虫作用的扫描电镜观察

目的 观察体外培养的卡氏肺孢子虫(Pc)经双氢青蒿素、青蒿琥酯作用后不同时相虫体表面结构变化。方法 将Pc接种入含HepG-2细胞的培养皿内并分别加入双氢青蒿素50μmol/L、青蒿琥酯100μmol/L、喷他脒15μmol/L、0.3％乙醇与空白对照。双氢青蒿素组于用药后1、2、4、8、12、16h作扫描电镜观察,后4组于用药后12h取样作扫描电镜观察。结果 双氢青蒿素作用2h后,Pc表面开始出现损伤,最初为表面绒毛脱落,残存绒毛肿胀呈球状,虫体体积增大。随着时间延长,虫体表面损伤明显,8h后表膜出现大小不等孔洞。青蒿琥酯组改变相同,但较轻微。结论 青蒿素衍生物可引起Pc虫体表膜损伤,通透性增加,功能障碍。     

卡氏肺孢子虫在八种细胞系上的增殖情况

目的　了解卡氏肺孢子虫在HepG 2 (人肝癌细胞 )等 8种细胞系的增殖状况 ,寻找适合卡氏肺孢子虫 (Pc)增殖的支持细胞系。方法　分别以 5× 10 5个虫体接种于 8种细胞系上。另采用HepG 2细胞 ,分别接种不同数量的Pc ,观察 7d ,比较Pc在不同细胞及不同接种数量的增殖情况。 结果　在HepG 2细胞上Pc滋养体可增殖 7倍左右 ,增殖高峰大多在第 4、5天。接种 5× 10 5个Pc ,包囊和滋养体均可较好增殖。结论　HepG 2细胞是较好的卡氏肺孢子虫体外培养支持细胞系 ,接种量以每孔 5× 10 5个Pc包囊为宜     

国产与进口白果内酯治疗大鼠卡氏肺孢子虫肺炎的比较研究

目的　比较国产和进口白果内酯治疗实验大鼠肺孢子虫肺炎的疗效。　方法　用Wistar大鼠建立卡氏肺孢子虫肺炎动物模型 ,分别用国产和进口的白果内酯 2 0mg/kg·d治疗实验大鼠 ,以免疫抑制大鼠和磺胺治疗大鼠为对照组 ,通过大鼠体重变化 ,肺重 /体重比值 ,肺印片的每视野包囊均数等指标考核疗效。　结果　国产和进口白果内酯治疗后两组大鼠的体重均增加 ,体重差分别为 (18.78± 16.82 ) g和 (17.85± 14 .86)g ,明显高于免疫抑制对照组 (2 .5 0±19.11)g(P <0 .0 5 ) ,两试验组间差异无显著性 (P >0 .0 5 ) ;肺重 /体重比值分别为 (1.2 7± 0 .5 2 )和 (1.3 0± 0 .62 ) ,低于免疫抑制对照组 (1.85± 0 .64 ) (P <0 .0 5 ) ,两试验组间差异无显著性 (P >0 .0 5 ) ;两试验组肺印片的每视野包囊均数分别为 (0 .96± 0 .3 6)和 (0 .69± 0 .19) ,低于对照组 (1.73± 0 .5 6) (P <0 .0 5 ) ,而国产白果内酯高于进口白果内酯组 (P <0 .0 5 )。结论　国产与进口白果内酯对大鼠肺孢子虫肺炎均有治疗作用 ,但国产白果内酯的疗效略差于进口白果内酯。     

卡氏肺孢子虫低死亡率SD大鼠动物模型的建立

目的建立低死亡率卡氏肺孢子虫肺炎(PCP) SD 大鼠动物模型. 方法将雌性SD大鼠随机分为实验组和对照组,实验组采用按体重定量皮下注射地塞米松免疫抑制的方法诱导建立PCP动物模型,对照组注射与地塞米松等体积的生理盐水.分别制作肺印片,经瑞-姬氏复合染色后,检查卡氏肺孢子虫包囊.制作肺组织病理切片,经HE染色后观察肺组织病理变化.制作感染大鼠肺组织超薄切片,透射电镜观察Pc包囊和滋养体. 结果用地塞米松诱导后,实验组SD大鼠死亡率为0,肺印片阳性率为76.7%(23/30).肺组织出现典型的病理变化,并可观察到Pc包囊.实验组SD大鼠体重下降明显,与对照组体重比较具有极显著性差异(P<0.01).电镜下可观察到Pc包囊和滋养体的形态结构. 结论采用按体重定量皮下注射地塞米松的方法可建立低死亡率PCP动物模型.     

Spontaneous pneumothorax in AIDS patients with recurrent Pneumocystis carinii pneumonia despite aerosolized pentamidine prophylaxis

Aerosolized pentamidine prophylaxis for Pneumocystis carinii pneumonia in patients with the acquired immunodeficiency syndrome (AIDS) may predispose these patients to recurrent apical Pneumocystis infection. Bullous changes and pulmonary cysts develop in the lung apices due to repeated episodes of inflammation and cytotoxic effects of HIV on pulmonary macrophages. These changes progress despite prophylaxis against recurrent Pneumocystis infection with aerosolized pentamidine, increasing the risk of spontaneous pneumothorax. Two cases are presented of bilateral pneumothoraces in patients with AIDS and recurrent P carinii pneumonia despite aerosolized pentamidine prophylaxis. Patients receiving aerosolized pentamidine prophylaxis for Pneumocystis pneumonia appear to have an increased risk of pneumothorax due to recurrent apical infections with P carinii.     

Intravenous or inhaled pentamidine for treating Pneumocystis carinii pneumonia in AIDS. A randomized trial

OBJECTIVE: To evaluate the efficacy and toxicity of aerosolized pentamidine and of reduced-dose intravenous pentamidine for the treatment of mild to moderate Pneumocystis carinii pneumonia in patients with the acquired immunodeficiency syndrome (AIDS). DESIGN: Randomized open study with serial pulmonary function testing and measurement of pentamidine concentrations in plasma and bronchoalveolar lavage fluid. PATIENTS: Of 44 men and 1 woman with a mild to moderate first episode of P. carinii pneumonia (Pao2 greater than or equal to 7.3 kPa [55 mm Hg]), 23 received aerosolized pentamidine and 22, intravenous pentamidine. INTERVENTIONS: Pentamidine isethionate, 600 mg by inhalation using a Respirgard II nebulizer (Marquest Medical Products, Inc., Englewood, Colorado) or 3 mg/kg body weight intravenously, administered once daily for 2 to 3 weeks. MEASUREMENTS AND MAIN RESULTS: The planned 60-patient study was stopped after 45 patients had been enrolled. The rates (aerosolized compared with intravenous pentamidine) of initial failure, early recrudescence of symptoms, and relapse were 12% and 19% (difference, 7%; 99% confidence interval [CI], - 23% to 37%; P = 0.67), 35% and 0% (difference, 35%; CI, 13% to 58%; P = 0.02), and 24% and 0% (difference, 24%; CI, 4% to 49%; P = 0.03). The rates (aerosolized compared with intravenous pentamidine) of major toxicity were 0% (0 of 17 patients) and 10% (2 of 21 patients) (difference 10%; CI, -1% to 29%; P = 0.24). The mean (+/- SD) pentamidine concentration in bronchoalveolar lavage fluid for patients receiving aerosolized pentamidine was 96.6 +/- 65.1 ng/mL compared with 14.4 +/- 17.7 ng/mL for patients receiving intravenous treatment. Trough concentrations of pentamidine in plasma increased from 0 to 25.4 +/- 16.4, 56.5 +/- 26.1, and 61.1 +/- 56.0 ng/mL at the end of weeks 1, 2, and 3 of intravenous therapy, respectively. CONCLUSIONS: The data suggest that reduced-dose intravenous pentamidine was more effective than aerosolized pentamidine for treating mild to moderate P. carinii pneumonia. Systemic absorption during aerosolized therapy was minimal; daily doses of intravenous pentamidine resulted in increased accumulation of pentamidine in plasma.     

Aerosolized pentamidine: effect on diagnosis and presentation of Pneumocystis carinii pneumonia

STUDY OBJECTIVE: To determine the effect of previous aerosolized pentamidine therapy on diagnosis and presentation of Pneumocystis carinii pneumonia. DESIGN: A retrospective study. SETTING: A tertiary care hospital. PATIENTS: Fifty-two consecutive patients with P. carinii pneumonia and underlying infection with the human immunodeficiency virus (HIV) who had bronchoscopy. Twenty-one patients who were on aerosolized pentamidine therapy served as the study group. Thirty-one patients who had not received the drug served as the control group. MEASUREMENTS AND MAIN RESULTS: The yield of bronchoalveolar lavage for P. carinii pneumonia was 62% for the study group and 100% for the control group (P less than 0.05). This lower yield was significant for the subset of patients having their first episode of P. carinii pneumonia. The yield of transbronchial biopsy was similar for both groups of patients (81% compared with 84%). The yield of bronchoscopy was not influenced by use of zidovudine. Review of lavage specimen slides suggested that there may be fewer organisms present in patients receiving aerosolized pentamidine. An atypical roentgenographic presentation of upper lobe predominant infiltrates was seen in 38% of the study patients and 7% of the control patients. In addition, pneumothoraces and cystic changes were also frequently seen in the study patients. Gallium scans, when done, were also atypical in the study group. Markers of the severity of disease, however, were similar in both groups. CONCLUSION: The yield of bronchoalveolar lavage for P. carinii pneumonia in HIV-infected patients is lower in patients receiving aerosolized pentamidine. Unusual roentgenographic presentations and atypical gallium scans are also found in this setting.     

Pneumothorax in AIDS

OBJECTIVE: To determine risk factors for the development of pneumothorax in patients with the acquired immunodeficiency syndrome (AIDS). DESIGN: Prospective cohort study. SETTING: Tertiary care center. PATIENTS: Of 1030 patients with AIDS who were followed at Memorial Sloan-Kettering Cancer Center between 1 January 1980 and 30 September 1989, 20 (2%) developed pneumothorax that was unrelated to trauma or a pulmonary procedure. RESULTS: Of 20 patients with AIDS who presented with pneumothorax, 19 had compelling evidence of concurrent Pneumocystis carinii pneumonia. Using bivariate analysis, patients receiving aerosol pentamidine prophylaxis (relative risk, 17.6) and those with a history of P. carinii pneumonia (relative risk, 14.5) were more likely to develop pneumothorax. By Mantel-Haenszel stratified analysis, aerosol pentamidine use was a statistically significant risk factor independent of a history of P. carinii pneumonia. The pneumothorax-related mortality rate was 10% and there was considerable morbidity. CONCLUSIONS: Patients with AIDS at the highest risk for developing pneumothorax are those with a history of P. carinii pneumonia who are receiving aerosol pentamidine prophylaxis but who nevertheless develop P. carinii pneumonia. The benefits of aerosol pentamidine prophylaxis in these patients far outweigh this risk. Pneumocystis carinii pneumonia should be considered as the most likely diagnosis in any patient with AIDS who develops a pneumothorax.     

Trimethoprim-sulfamethoxazole compared with pentamidine for treatment of Pneumocystis carinii pneumonia in the acquired immunodeficiency syndrome. A prospective, noncrossover study

STUDY OBJECTIVE: To ascertain the efficacy and toxicity of trimethoprim-sulfamethoxazole or pentamidine when either is given alone during the entire treatment period for Pneumocystis carinii pneumonia in patients with the acquired immunodeficiency syndrome (AIDS). DESIGN: Prospective, randomized, noncrossover comparison of trimethoprim-sulfamethoxazole with pentamidine. Trimethoprim-sulfamethoxazole dosage was adjusted to maintain serum trimethoprim at 5 to 8 micrograms/mL. Pentamidine dosage was reduced by 30% to 50% for an absolute rise in serum creatinine of more than 88 mumol/L (1 mg/dL). SETTING: Tertiary care hospital and AIDS clinic. Patients: Thirty-six patients were treated with trimethoprim-sulfamethoxazole and 34 with pentamidine. Pretreatment clinical features and laboratory test results were similar in the two groups. MEASUREMENTS AND MAIN RESULTS: Thirty-six recipients of trimethoprim-sulfamethoxazole and 33 recipients of pentamidine completed therapy without crossover. Trimethoprim-sulfamethoxazole caused a rash (44%) and anemia (39%) more frequently (P less than or equal to 0.03, whereas pentamidine caused nephrotoxicity (64%), hypotension (27%), or hypoglycemia (21%) more frequently (P less than or equal to 0.01). The (A - a)DO2 improved by greater than 1.3 kPa (10 mmHg) 8 days earlier for trimethoprim-sulfamethoxazole recipients (95% CI for the difference in response, -1 to 17; P = 0.04). Thirty-one (86%) patients treated with trimethoprim-sulfamethoxazole and 20 (61%) with pentamidine survived and were without respiratory support at completion of treatment (95% CI for the difference in response, 5% to 45%; P = 0.03). CONCLUSIONS: For most patients with AIDS and P. carinii pneumonia, successful treatment with a single agent is possible. Toxicity associated with the two standard treatments is rarely life-threatening and may be diminished if the trimethoprim-sulfamethoxazole dosage is modified by pharmacokinetic monitoring and the pentamidine dosage is reduced for nephrotoxicity. Oxygenation improved more quickly and survival was better with trimethoprim-sulfamethoxazole.     

Effect of aerosolized pentamidine prophylaxis on the diagnosis of Pneumocystis carinii pneumonia by induced sputum examination in patients infected with the human immunodeficiency virus

This study assessed the effect of aerosolized pentamidine prophylaxis on the clinical presentation and diagnostic sensitivity of induced sputum examination for Pneumocystis carinii pneumonia. Between January 1, 1988 and October 27, 1990, 348 induced sputum examinations were performed as the initial diagnostic procedure for P. carinii pneumonia in patients infected with the human immunodeficiency virus (HIV). Medical records were reviewed for all induced sputum examinations, and the study group consisted of patients who either had not received prophylactic therapy (n = 193) or had received aerosolized pentamidine prophylaxis (n = 126). A total of 29 induced sputum examinations in patients receiving either other prophylactic regimens or ongoing therapy for previously documented P. carinii pneumonia were excluded from the study group. A total of 72 consecutive episodes of P. carinii pneumonia were subsequently documented by induced sputum examination (n = 54), bronchoalveolar lavage (n = 16), thoracocentesis (n = 1), or autopsy (n = 1). A total of 44 episodes occurred in patients who had not received antipneumocystis prophylaxis, and 28 episodes occurred in patients who had received aerosolized pentamidine. Of patients capable of producing a sputum specimen for analysis, induced sputum examination had a significantly lower diagnostic yield of 64.3% in patients who had received aerosolized pentamidine prophylaxis compared with 92.3% in patients who did not receive prophylaxis (p less than 0.02, Fisher's exact test). When the data were analyzed on an intention to treat basis, although there was a trend suggesting a lower overall yield in the aerosolized pentamidine patients, the difference was not statistically significant (64.3 versus 81.8%, p = 0.17, Fisher's exact test).(ABSTRACT TRUNCATED AT 250 WORDS)     

Pneumocystis hepatitis and choroiditis despite successful aerosolized pentamidine pulmonary prophylaxis

A patient who developed Pneumocystis carinii hepatitis and choroiditis despite receiving prophylactic pentamidine therapy by aerosol is described. Liver biopsy showed histology typical of Pneumocystis hepatitis, but his respiratory status was stable and his lungs were free of P carinii organisms on BAL. Thus, inhaled pentamidine prophylaxis did not prevent extrapulmonary pneumocystosis. Patients receiving pentamidine prophylaxis with unexplained symptoms should undergo investigation for possible extrapulmonary P carinii infection.     

丝裂原蛋白激酶两种抑制剂对弓形虫侵入宿主细胞影响的差异

目的观察两种丝裂原蛋白激酶(mitogen activated protein kinases,MAPK)抑制剂PD98059及U0126对刚地弓形虫侵入宿主细胞的影响,探讨其对弓形虫速殖子侵入宿主细胞信号转导途径的不同阻断效应。方法丝裂原蛋白激酶抑制剂PD98059或U0126分别在不同时间及不同剂量作用于速殖子-宿主细胞培养系统,用流式细胞仪(FCM)检测宿主细胞感染速殖子的差异。结果流式细胞仪检测加入U01261μmol/L、10μmol/L和100μmol/L的细胞培养孔的细胞感染弓形虫速殖子的量分别比对照组平均降低了26.10%(P<0.01),66.42%(P<0.01)和70.39%(P<0.01)。而加入PD980591μmol/L、10μmol/L和100μmol/L的分别比对照组平均降低了25.45%(P<0.01),53.01%(P<0.01)和64.70%(P<0.01)。实验中发现100μmol/L U0126作用培养细胞9h的时候,可导致HL-60细胞出现部分聚集成团、漂浮的中毒现象。结论U0126和PD98059均可明显抑制弓形虫速殖子侵入宿主细胞,但其差异无显著性,其机制有待进一步探索。