卡氏肺孢子虫肺炎动物模型的实验观察

目的：建立卡氏肺孢子虫肺炎(PCP)动物模型并观察其病理变化。方法：将Wistar大鼠随机分为A、B实验组(每组5只)和对照组(4只)。A、B实验组用地塞米松加四环素免疫抑制，对照组只用四环素。A组和对照组喂低蛋白质食物，B组喂高蛋白质食物。结果：A、B实验组病死鼠肺印片均查到PC包囊；A、B实验组病理组织学形态、感染数和死亡数无明显差异，对照组健康存活。结论：免疫功能低下时可致PCP，此模型的建立有助于PCP的研究。     

Wistar大鼠卡氏肺孢子虫肺炎动物模型的建立

卡氏肺孢子虫研究材料的主要宿主来源是免疫抑制大鼠。Wistar大鼠10只,腹腔注射醋酸可的松25mg/只,2次/周,低蛋白饮食。8周后检查大鼠卡氏肺孢子虫。结果表明,全部大鼠肺印片和肺悬液涂片中均可检出卡氏肺孢子虫。     

卡氏肺孢子菌肺炎动物模型建立及其病原体的超微结构观察

目的建立SD大鼠的卡氏肺孢子菌肺炎(PCP)的实验动物模型,观察肺组织病理变化特点和卡氏肺孢子菌(Pc)的超微结构特征,探讨其致病机制。方法地塞米松皮下注射诱导SD大鼠PCP,肺印片吉姆萨染色检测Pc滋养体,GMS染色检测Pc包囊,肺组织切片HE染色观察肺组织病理变化,透射电镜观察Pc超微结构。结果地塞米松诱导6周后,大鼠肺印片可见少许Pc滋养体和少许包囊,第9周肺泡腔内可见大量滋养体和包囊;PCP肺泡腔变小,肺泡上皮增生,间隔增宽,肺间质内有以淋巴细胞、嗜酸性粒细胞为主的细胞浸润并伴炎性物质渗出,并随病程时间延长炎症进行性加重;透射电镜显示,Pc黏附在肺Ⅰ型上皮细胞,滋养体形态多样,表面有管状突出和伪足样结构,内部含有丰富的线粒体、内质网、管状小体、囊内小体、高密度特殊颗粒等超微结构;包囊表面有皱褶但未见管状突起,囊内有数个囊内小体,具有滋养体样特征。结论 Pc对肺Ⅰ型上皮细胞的黏附和过度繁殖增生以及刺激肺间质大量炎性细胞浸润和炎性物质渗出是引发PCP不可逆肺损伤的重要因素。     

地塞米松抑制烟曲霉菌暴露的支气管哮喘大鼠肺组织IL-33的表达

 目的  研究烟曲霉菌暴露对支气管哮喘大鼠肺组织IL-33水平的影响及地塞米松的作用。方法  将24只Wistar大鼠随机分为A组(正常对照组)、B组(哮喘组)、C组(烟曲霉菌暴露组)、D组(地塞米松干预组),每组6只。B、C、D组均以卵白蛋白(ovalbumin,OVA)致敏、激发的方法构建哮喘模型;C、D组在哮喘模型构建成功后,给予烟曲霉菌孢子鼻腔滴入;D组在OVA激发阶段给予地塞米松干预;A组则予等量生理盐水致敏、激发、滴鼻作为对照。通过气道高反应性检测、嗜酸性粒细胞百分比及血清IgE水平确定哮喘模型的建立,ELISA法及qRT-PCR法检测肺组织IL-33的表达,大鼠肺组织和全血于马铃薯葡萄糖琼脂培养基(potato dextrose agar,PDA)上培养24 h后观察烟曲霉菌菌落生长情况。结果  B、C组与A组比较,肺泡灌洗液中嗜酸性粒细胞百分比、血清IgE水平及肺组织IL 33的表达均明显升高(P<0.05),且C组比B组升高更明显。各组间气道反应性指标sRaw变化值差异不明显。D组与C组比较,嗜酸性粒细胞百分比及肺组织IL-33的表达均明显降低(P<0.05),血清IgE水平降低不显著,但肺组织烟曲霉菌培养阳性率高于C组。结论  糖皮质激素(地塞米松)能够抑制烟曲霉菌暴露的支气管哮喘大鼠肺组织IL-33的表达,但增加了气道真菌定植的风险,针对IL-33的阻断治疗有待进一步研究。     

卡氏肺孢子虫病动物模型建立和虫体形态观察

采用醋酸可的松诱发Wistar大鼠、BALB/C小鼠和家兔感染卡氏肺孢子虫,比较不同动物对卡氏肺孢子虫的易感性。经姬氏染色法、哥氏银染色法等染色后进行虫体形态观察,结果显示,Wistar大鼠卡氏肺孢子虫惑染率最高(100%),虫体量多。不同给药方式对Wistar大鼠卡氏肺孢子虫的感染率无显著性意义。各种染色法可显示肺组织印片或切片的滋养体或包囊。姬氏染色法是显示印片中虫体的可靠方法。改良哥氏银染色法和快速焦油紫法是显示肺组织石蜡切片中包囊的可靠方法。伦氏荧光桃红——肼黄法能良好地显示切片中的滋养体和囊内小体。     

卡氏肺孢子虫肺炎动物模型建立及其检测方法

用皮下注射醋酸可的松法诱导大鼠卡氏肺孢子虫肺炎(PCP)模型,成功率20％。对PCP模型大鼠(n=6)进行诊断采样,结果:肺病理学检查卡氏肺孢子虫(PC)检出率为100％(6/6)。用经支气管肺活检(TBLB)法临床诊断1例早期PCP。支气管肺泡灌洗液(BALF)涂片和肺组织印片的姬姆萨染色,可见PC囊内小体及滋养体。改进的哥氏银染适用于肺组织切片,包囊壁浓染呈黑色,易于识别。对于免疫功能低下怀疑为PCP患者,应尽早作TBLB和(或)BAL检查,可以达到早期诊断。     

氨苯砜和蒿甲醚治疗大鼠卡氏肺孢子虫肺炎的疗效研究

目的研究氨苯砜和蒿甲醚对实验大鼠卡氏肺孢子虫肺炎的疗效。方法将wistar大鼠随机分为A、B实验组,以地塞米松皮下注射,诱发建立卡氏肺孢子虫肺炎动物模型。A组实验鼠5只,用氨苯砜治疗,50mg／只·次,灌服,2次/d,连用10d;B组实验鼠6只,用蒿甲醚治疗,20mg／只·次,肌注,1次/d,连用5d,首剂加倍。A、B组分别有1只为不治疗对照鼠。结果A组实验鼠在治疗后第12d基本恢复健康,对照鼠在第3d死亡;B组实验鼠在治疗后第8d基本恢复健康,对照鼠在第2d死亡。结论氨苯砜和蒿甲醚对卡氏肺孢子虫肺炎均有疗效。     

瑞香素治疗大鼠肺孢子虫肺炎的实验研究

目的观察瑞香素对大鼠肺孢子虫肺炎(PCP)的治疗效果。方法皮下注射醋酸泼尼松龙建立PCP大鼠模型。将受试大鼠随机分为正常对照组(A)、未治疗对照组(B)、TMP-SMZ对照组(C)、瑞香素治疗组(D)和青蒿素对照组(E)5组。各组从第7周开始灌服相应药物5d,停药后观察1周,用肺质量/体质量比和肺组织虫荷量考核疗效。结果PCP大鼠6周内的病死率为16.7%;满6周者抽样进行病原、病理检查证实均已成功诱导PCP。A、B、C、D、E组受试大鼠的平均肺质量/体质量比分别为0.75、1.05、0.77、0.77和0.85,B~E4组受试大鼠肺组织虫荷量分别为1490、74、339和219个/mg。统计学分析显示,经瑞香素治疗的大鼠平均肺质量/体质量比、肺虫荷量均明显低于未治疗对照组(分别为P<0.05和P<0.01),效果虽不如TMP-SMZ,但与青蒿素疗效相似(P>0.05)。结论瑞香素对大鼠PCP有显著疗效,效果与青蒿素接近。     

不同海拔大鼠肺气肿模型肺组织caspase-3水平比较

目的 观察不同海拔肺气肿大鼠肺组织caspase-3表达水平,评估低氧对肺组织凋亡的影响.方法 40只Wistar 大鼠随机分为对照组（A组,6只）、中海拔盐水组（B组,6只）、中海拔肺气肿组（C组,8只）、高海拔盐水组（D组,10只）、高海拔肺气肿组（E组,10只）.A、B、C3组大鼠饲养于西宁,海拔2260m.D组和E组饲养于低压氧舱,模拟海拔5500m.B组和D组腹腔注射生理盐水、C组和E组腹腔注射CSE,饲养4周后,采集大鼠右肺组织,免疫组化法检测肺组织caspase-3的表达.结果 与A、B、D3组比较,C组和E组肺实质破坏、部分肺泡破裂及肺泡腔扩大.5组间caspase-3细胞阳性率比较有统计学差异（P＜0.001）.D组高于B组及A组（P ＜0.001）,E组高于C组（P＜0.001）,C组高于B组（P＜0.001）,E组高于D组（P＜0.001）.结论 高原低氧加重了肺气肿大鼠的凋亡表达.     

艾滋病合并卡氏肺孢子菌肺炎一例

AIDS进展期合并的肺部感染中42％为卡氏肺孢子菌肺炎（PCP），PCP依据临床表现、胸部X线征象及痰涂片找到卡氏肺孢子菌或经聚合酶链反应（PCR）阳性而确诊，SMZ—TMP是治疗和预防的首选。本文以一例进行性呼吸困难为首发症状的AIDS合并卡氏肺孢子菌肺炎患者为例探讨AIDS合并PCP感染患者的诊治。     

两种单肺通气方式对大鼠肺AQP-5表达影响的对比研究

目的研究两种单肺通气（OLV）方式下大鼠肺组织水通道蛋白-5（AQP-5）表达的变化。方法建立两种OLV大鼠模型：夹闭左侧肺门OLV组（A组）,过深插管OLV组（B组）。按不同通气时间分亚组A1、A2、A3和B1、B2、B3。设双对照组：双肺通气组（C组）,同样分亚组C1、C2、C3;空白对照（D组）。免疫印记法检测肺AQP-5的表达。结果 A、B、C组AQP-5的表达均较D组减少（P〈0.05）;各亚组左右两侧肺AQP-5的表达差异无统计学意义（P〉0.05）;A、B、C组组内各亚组比较差异均有统计学意义（P〈0.05）;A1、B1、C1组间比较差异均无统计学意义（P〉0.05）;A2、B2、C2组间比较,A2比B2和C2均有减少（P〈0.05）,B2与C2差异无统计学意义（P〉0.05）;A3、B3、C3组间比较,A3和B3均较C3组减少（P〈0.05）,A3比B3减少（P〈0.05）。结论机械通气可致大鼠AQP-5表达下调,与时间相关,夹闭法OLV和插管过深法OLV对AQP-5表达的影响超过双肺通气,前者更明显。     

重组腺病毒Ad-HGF对烟熏大鼠肺气肿肺组织细胞凋亡的抑制作用

目的通过构建大鼠肺气肿模型,观察携带肝细胞生长因子基因的重组腺病毒（Ad-HGF）对肺组织细胞凋亡的抑制作用。方法40只Wistar大鼠随机分为4组：正常对照组（A组）、肺气肿组（B组）、肺气肿＋Ad～HGF组（C组）和肺气肿＋Ad—GFP组（1）组）;采用烟熏法建立大鼠肺气肿模型。造模成功后,A组正常饲养,B组给予0．5ml生理盐水灌注治疗,C组给予1×10^9 pfu Ad-HGF,D组给予1×10^9 pfuAd-GFP：干预14天后处死各组动物,取D组大鼠部分肺组织作常规冰冻切片,荧光显微镜下观察目的基因的表达;取A、B、C5-组大鼠肺组织制作病理切片,观察并计算平均肺泡面积（MAA）、平均内衬间隔（MLI）、平均肺泡数（MAN）;采用脱氧核糖核酸末端转移酶介导的dUTP缺口末端标记（TUNEL）技术对大鼠肺实质凋亡细胞的阳性细胞率（AI）进行检测。结果与A组相比,B组、C组大鼠的肺组织都存在不同程度的肺气肿病理改变;Ad—HGF治疗纽MAA和MLI均小于模型组、MAN大于模型组,差异有统计学意义（P〈O．05）;Ad—HGF治疗组肺组织细胞凋亡指数（AI）低于模型组,差异有统计学意义（P〈0．05）。结论重组腺病毒Ad—HGF对烟熏大鼠肺气肿肺组织细胞凋亡有抑制作用。     

地塞米松对大鼠肺缺血再灌注后eNOS表达的影响

目的：通过观察地塞米松对大鼠肺缺血再灌注后内皮型一氧化氮合酶（eNOS）的影响探讨其对缺血再灌注肺的保护机制。方法：40只健康SD大鼠,雌雄不拘,随机分为5组：假手术组（S）、单纯缺血组（I）、缺血再灌注组（IR）、小剂量地塞米松干预组（D1）及大剂量地塞米松干预组（D2）,每组8只。根据Eppinger模型制作SD大鼠单侧肺IRI动物模型,Western-blot法检测eNOS蛋白质浓度,HE染色观察组织病理学改变。结果：与其它组比较,IR组肺组织中eNOS表达明显下降（P〈0.01）,地塞米松干预后,eNOS表达明显增高,以D2组明显。病理切片示IR组肺组织结构损害分级显著重于其它组,地塞米松干预后肺组织结构损害分级下降,以D2组明显。结论：eNOS在正常肺组织中表达较多,IR可使eNOS表达明显下降,地塞米松可使eNOS表达增高,尤以大剂量时明显,地塞米松通过上调eNOS表达减轻缺血再灌注肺组织损伤。     

Relationship between the burden of pneumocystis carinii, the inflammatory reaction and lung injury in pneumocystis carinii pneumonia

Objective To study the relationship between the burden of Pneumocystis carinii (P. carinii) and the inflammatory reaction and biochemical markers in bronchoalveolar lavage fluids (BALF)in a rat model of P. carinii pneumonia (PCP). Methods Clean grade 50 male Sprague-Dawley rats were immunosuppressed by a subcutaneous injection of 25mg cortisone acetate twice a week for 8-12 weeks; the PCP model was successfully induced in 14 rats. The inflammatory reaction and biochemical markers of the activity of lactate dehydrogenase (LDH), alkaline phosphatase (AL P) and type Ⅳ collagenase (matrix metalloproteinases, MMP-2, MMP-9) as well a s the values of total protein (TP) and albumin (ALB) in BALF between the mild burden group of P. carinii (involved alveoli &lt;25% per 100 alveoli, G roup A) and the moderate to severe burden group (involved alveoli ≥25% per 100 alveoli, Group B) were measured. The other six clean grade SD rats served as no rmal control group (Group C).Results The total white cell count in BALF was higher in Group B ［(6.8±1.7)×10(6)/L ］ than in Group A ［(3.8±1.2)×10(6)/L］ (P&lt;0.01); however, there were no differences in white cell differentiation. Assays of biochemical marke rs showed that ALB in BALF in Group B (0.893±0.469 g/L) was increased in com parison with Group A (0.262±0.169 g/L); it was only 0.026±0.021 g /L i n Group C. The contents of TP and activities of LDH were higher in Group B (TP 1.756±0.706 g/L, LDH 2580±550 U/L) than in Group A (TP 0.784±0.553 g/L, LDH 1410±620 U/L); the values of TP and LDH were 0.063±0.020 g/L an d 370±250 U/L respectively in Group C. The activity of Type Ⅳ collagenase, including MMP-2 and MMP-9, was higher in Group B than in Group A (P&lt;0.0 1) (MMP-2: 1102±169 grey value vs 459±274 grey value; MMP-9: 1218±257 grey value vs 449±225 grey value). There was no activity of Type Ⅳ collagenase in BALF of Group C. No statistically significant difference was observed in ALP between the groups B and A. Conclusions These results indicate that there is a significant correlation between the burde n of P. carinii in lung tissues and the inflammatory reaction as well as bi ochemical markers of the resultant activity of lung injury.     

卡氏肺孢子虫小鼠动物模型的研究

目的：建立卡氏肺孢子虫小鼠动物模型。方法：昆明小鼠１６只，每次皮下注射醋酸可的松１～２ｍｇ，每周２次，诱发小鼠卡氏肺孢子虫肺炎模型。结果：用药５周肺印片即可检出卡氏肺孢子虫包囊，８周后大部分小鼠可检出，总检出率为６８．７５％。姬氏染色肺印片可查见成熟包囊、未成熟包囊，形态典型，是显示肺印片中包囊的可靠方法。ＧＭＳ染色是显示肺组织石蜡切片中包囊的可靠方法。结论：提示小鼠可能是建立卡氏肺孢子虫动物模型的理想动物     

地塞米松诱发大鼠卡氏肺孢子虫肺炎实验研究

目的 在地塞米松磷酸钠诱导下建立大鼠卡氏肺孢子虫肺炎的动物模型。方法 将40只雌性SD大鼠随机分为实验组和对照组,前者皮下注射地塞米松磷酸钠,1mg/次,2次/w;后者不做任何处理。观察两组大鼠的发病情况,并每隔3周两组各取5只动物进行病原学检查。结果 实验组大鼠第6周开始发病,肺印片、支气管肺泡灌洗液沉渣中查到卡氏肺孢子虫包囊及滋养体;对照组大鼠无异常表现,病原学检查阴性。结论 在地塞米松磷酸钠诱导下可成功建立大鼠卡氏肺孢子虫肺炎动物模型。     

Genotypes and polymorphisms of mutant CCR5-△32,CCR2-64I and SDF1-3’A HIV- 1 resistance alleles in indigenous Han Chinese

Objective To understand the interaction between surfactant proteins and pneumocys tis carinii pneumonia (PCP), and the impact of corticosteriods on surfactant proteins.Methods We established rat models of PCP and bacterial pneumonia induced by subcutaneous injection of 25mg cortisone acetate. At 8-12 wk, the bronchoalveolar lavage f luid (BALF) of rats was collected. Total nucleated cells of BALF were counted a nd differentiated, and the concentrations of surfactant protein A (SP- A) and su rfactant protein D (SP- D) were measured by immunoblotting assay. The rats were divided into three immunosuppressive groups and a normal control group. Group Ⅰ, normal control (n=6), consisted of healthy SD rats; group Ⅱ, negative cont rol (n=6), consisted of rats with cortisone acetate injection for over 8 wk with out lung infection; group Ⅲ, bacterial pneumonia (n=11), rats were injected wit h cortisone acetate over 8 wk that resulted in bacterial pneumonia without other pathogens isolated; and group Ⅳ, PCP (n=14), rats with injected cortisone acet ate for 8-12 wk and developed PCP without other pathogens isolated. Results Our results indicated that the total cell count in BALF in the negative control group was lower than that in the normal control group (P&lt;0.001). During P CP infection, the total cell count and the percentage of polymorphonuclearcytes (PMNs) in BALF were signi ficantly increased (P&lt;0.01), but were lower than those in the bacterial pne umonia group. The concentration of SP- A of BALF in PCP (45.1±22.1 μg/ml) was significantly increased in comparison with that in the negative control ( 16.2±9.9 μg/ml, P&lt;0.05) and bacterial pneumonia groups (6.2±5.6 μ g/ml, P&lt;0.001). We also found that the relative content of SP - D was signi ficantly higher in PCP (24 249±4780 grey values) than that in the negative control (13 384±2887 grey values, P&lt;0.001) and that in bacterial pne u monia (11 989±2750 grey values, P&lt;0.001). SP- A and SP- D were a lso higher in the moderate to heavy group of PCP than those seen in the mild group (P&lt;0.01, P&lt;0.001). SP- A and SP- D were higher in the negative contr ol group than those in the normal control group, but there was no significant di fference between the 2 groups. Conclusion These results suggest that the concentrations of SP- A and SP- D in BALF are inc reased by pneumocystis carinii specific stimulation, but the alteration is n ot related to the corticosteriod usage.