Protective effect of DNA vaccine with the gene encoding 55kDa antigen fragment against Pneumocystis carinii in mice

Objective:To evaluate the protective effect of DNA vaccine with the gene encoding 55kDa antigen fragment of Pneumocystis carinii(P.carina) against P.carina in mice.Methods:The fragment of the antigen within p55(p55-582) was cloned.Then recombinant plasmid was constructed based on the eukaryotic expression vector pcDNA3.1(+).BALB/c mice were used as experimental models to examine the immunogenicity of pcDNA3.1(+)-p55-582.ELBA and RTPCR were used to evaluate the role of this kind of DNA vaccine.Results:The results of western blot indicated that the recombinant DNA[pcDNA3.1(+)-p55-582]could be expressed correctly and had antigenicity in transfected COS-7 cells.ELBA and RT-PCR showed that pcDNA3.1(+)- p55-582 elicited antibody production,stimulated lymphocyte proliferation and provided partial protection by reducing the P.carina burden.Conclusions:The data demonstrate that pcDNA3.1(+)-p55-582 might be potent vaccination that can afford the partial protection for the immunized animals.

A Study on Using Improved Methenamine-silver Stain to Diagnosis of Pneumocystis carinii

Lung smears of mice and lung sections of rats or human case with Pneumocystis cariniiinfection were stained using the Grocott's modification method of Gomori's methenamine-silver nitratetechnic, in which 5% sodium periodate and 5% chromic acid were used as oxidant respectively. Theoxidation time for the mouse lung smears was 5,15,60 minutes and the oxidation temperature was 20℃.The time of silver impregnation was 90 minutcs and the temperature was 60℃ for the all smearo. Whenthe oxidation time was under 15 minutes. Pneumocystis cariniic cysts showed light or dark brown, and theparenthesis-like structure could clearly be found in part of the cysts. However, if the time of oxidationWas longer, the cysts showed black and secmed to have damaged. In the same batch of the mouse lungsmears oxidated for 5 minutes, the samiples oxidated by sodium periodate showed more the cysts with theparen thesis-like structure than those oxidated by chromic acid.In the rat or patient's lung sectionsoxidated by. sodium periodate, this structure could also be found. The result of the experiment showsthat sodium periodate as an oxidant in the subsequent step of the the silver impregnation is preferable tochromic acid. And then,it is useful to clinical practice that the step of sodium bisulfate can be omittedin the study.

Analysis of 8 chronic kidney disease patients complicated with Pneumocystis carinii pneumonia

Objective To study the clinical characteristics and outcome of Pneumocystis carinii pneumonia(PCP) in patients with chronic kidney diseases.Methods Clinical data of 8 cases of chronic kidney diseases complicated with PCP(excluding renal transplant patients) were examined retrospectively.Results The most common presenting symptoms at admission were fever(100%),cough without or with a little sputum(87.5%),and exertional dyspnea(75%).Beside these,they complained of chest tightness,fatigue,sweating and chills.Six patients(75%) presented with hypoxemia were diagnosed with type 1 respiratory failure during the course of illness.The most common CT feature was bilateral patchy areas of ground-glass opacities.Five patients had peripheral blood lymphocyte count less than 1 ×109/L.Four patients had CD4 cell count less than 200/mm3.Serum LDH level was elevated in 5 patients(582±222.55).Among the 8 patients,2 patients died within 20 days of PCP diagnosis.Conclusion Pneumocystis carinii pneumonia is an opportunistic and serious complication in chronic kidney disease patients treated with immunosuppressants.The disease progression is fast and patients with respiratory failure have a high mortality rate.Early diagnosis and appropriate treatment are important for better prognosis.

Clinical Analysis of 15 Cases of Non-Hodgkin Lymphoma Complicated with Pneumocystis carinii Pneumonia Treated with R-CHOP Regimen

Objective:To investigate the clinical features of R-CHOP regimen in the treatment of non-Hodgkin^lymphoma with Pneumocystis carinii pneumonia(PCP)in order to improve the understanding of PCP and the side effects of Rituxan.Methods:A retrospective analysis of 90 patients with non-Hodgkin’s lymphoma treated with R-CHOP chemotherapy in our hospital from November 2015 to November 2020,of which 15(16.7%)patients,combined with PCP clinical data,including clinical symptoms,physical signs,chest imaging examination and treatment data were used for to analysis and summarization.Results:The clinical features of R-CHOP chemotherapy combined with PCP were fever,cough,and sputum.Some patients had fewer clinical symptoms.Common imaging manifestations were double lung membrane glass shadow,patchy shadow,and flocculent shadow.It can occur in all clinical stages,and the incidence of late stage is high,and there is no clear correlation with bone marrow suppression.Pneumocystis was found in 2 cases of sputum,and the rest of the patients were clinically diagnosed.The main therapeutic drugs are sulfamethoxazole(8/15),compound sulfamethoxazole(6/15),clindamycin(1/15,sulfa drug allergy),and adrenal cortex hormones(4/15).Fourteen cases were cured and 1 case died.Conclusion:The incidence of R-CHOP in advanced non-Hodgkin^lymphoma of PCP is high.Patients with clinical use of R-CHOP chemotherapy will encounter fever,cough,chest computed tomography(CT)film glass shadow,and diffuse patch shadow.Patients should be alert to the possibility of PCP and take sulfonamides as soon as possible for medical treatment.

Pentamidine in Pneumocystis jirovecii prophylaxis in heart transplant recipients

Despite advances in transplantation techniques and the quality of post-transplantation care, opportunistic infections remain an important cause of complications. Pneumocystis jirovecii(P. jirovecii) is an opportunistic organism, represents an important cause of infections in heart transplantation patients. Almost 2% to 10% of patients undergoing cardiac transplantation have Pneumocystis pneumonia. Prophylaxis is essential after surgery. Various prophylaxis regimes had been defined in past and have different advantages. Trimethoprim/sulfamethoxazole(TMP/SMX) has a key role in prophylaxis against P. jirovecii. Generally, although TMP/SMX is well tolerated, serious side effects have also been reported during its use. Pentamidine is an alternative prophylaxis agent when TMP/SMX cannot be tolerated by the patient. Structurally, pentamidine is an aromatic diamidine compound with antiprotozoal activity. Since it is not effectively absorbed from the gastrointestinal tract, it is frequently administered via the intravenous route. Pentamidine can alternatively be administered through inhalation at a monthly dose in heart transplant recipients. Although, the efficiency and safety of this drug is well studied in other types of solid organ transplantations, there are only few data about pentamidine usage in heart transplantation. We sought to evaluate evidence-based assessment of the use of pentamidine against P. jirovecii after heart transplantation.

艾滋病合并耶氏肺孢子菌肺炎和猴痘1例报道

猴痘病毒相关肺部感染较罕见,本文报道1例艾滋病合并耶氏肺孢子菌肺炎(pneumocystis jirovecii pneumonia,PJP)同时合并猴痘的病例。该病例系沈阳市第六人民医院2023年10月收治的40岁男性患者,入院前已确诊HIV阳性,周身皮疹伴发热2个月,干咳2周。皮疹局部可见脓性渗出及少量渗血,周围绕以红晕。CD4^(+)T淋巴细胞6.0×10^(6)/L,猴痘病毒核酸检测阳性。肺部影像学检查显示两肺呈弥漫性分布磨玻璃影,肺内见多发大小不等实性结节灶,边缘可见晕征、毛刺等。肺结节穿刺活检,组织病原微生物检测为猴痘病毒及耶氏肺孢子菌阳性。临床给予抗感染、抗真菌及复方新诺明等治疗,患者发热、呼吸困难持续加重,最终死亡。本文通过复习有关资料,深入探讨了艾滋病合并PJP和猴痘时的临床特点及肺部影像学表现,以期提升临床认识。

以合并肺孢子菌肺炎为例探讨新型冠状病毒感染的鉴别诊断及中西医治疗

随着疫情防控形势的调整,新型冠状病毒感染(简称新冠感染)与其他肺部感染性疾病的鉴别诊断尤显重要。肺孢子菌肺炎属真菌感染性肺炎,与新冠病毒同时感染时,其症状和肺部影像学表现常难以区分,易被误诊误治。本篇报道的2例肺孢子菌肺炎合并危重型新冠感染的艾滋病患者,在西医常规治疗的基础上,运用中医清肺解毒、益气养阴等治法,祛邪扶正,帮助减轻患者的症状,延缓病情的进展,促进核酸早日转阴。

肾移植术后并发卡氏肺孢子菌肺炎的治疗方案选择

目的:探讨肾移植术后并发卡氏肺孢子菌肺炎(PCP)患者用卡泊芬净治疗PCP感染的疗效。方法:将125例肾移植后并发PCP患者分为磺胺组(42例)、卡泊芬净组(37例)和磺胺+卡泊芬净组(46例),比较3组临床特征,分析各组治疗后患者体温恢复正常时间、卡氏肺孢子菌(PC)核酸转阴时间和治疗结局。结果:治疗前,3组患者年龄、性别、发热症状及实验室检测(G实验和乳酸脱氢酶)方面比较,差异无明显统计学意义(P均>0.05)。治疗后,磺胺组、卡泊芬净组、磺胺+卡泊芬净组体温恢复正常时间分别为(7.9±1.4)d,(8.1±1.7)d和(7.7±1.5)d,组间比较,差异无明显统计学意义(P>0.05);PC核酸转阴时间分别为(13.4±1.6)d、(16.3±2.9)d、(13.9±2.5)d;卡泊芬净组PC核酸转阴时间明显长于磺胺组、磺胺+卡泊芬净组(P均<0.05),磺胺组和磺胺+卡泊芬净组比较,差异无明显统计学意义(P>0.05);3组间的治疗结局比较,差异无明显统计学意义(P均>0.05)。结论:卡泊芬净单药治疗PCP能获得肯定的疗效,包括体温恢复正常、PC核酸转阴和治愈出院。卡泊芬净对于肾移植术后PCP患者的治疗是一个较好的选择。

卡泊芬净辅助治疗对HIV感染合并肺孢子菌肺炎患者CD4^(+)/CD8^(+)T淋巴细胞比值和血氧指标的影响

目的研究卡泊芬净辅助治疗对艾滋病毒(HIV)感染合并肺孢子菌肺炎(PCP)患者CD4^(+)/CD8^(+)T淋巴细胞比值及血氧指标的影响。方法前瞻性将2017年1月至2022年1月间汉中市中心医院收治的62例HIV感染合并PCP患者采用随机数表法分为观察组和对照组,两组31例。对照组患者应用复方磺胺甲恶唑治疗,观察组患者在此基础上加用卡泊芬净治疗,1周为一个疗程,两组均连续治疗3周。分别于治疗前后采集两组患者外周静脉血,采用酶联免疫吸附法(ELISA)检测血清乳酸脱氢酶(LDH)、1→3-β-D葡聚糖(BDG)以及涎液化糖链抗原-6(KL-6)水平,流式细胞仪(贝克曼库尔特)检测CD4^(+)、CD8^(+)水平,血气分析仪检测动脉血氧分压(PaO_(2))、血氧饱和度(SaO_(2));参照《侵袭性肺部真菌感染的诊断标准与治疗原则草案》评估两组患者的治疗效果,并统计两组患者的不良反应发生情况。结果治疗后,两组患者的LDH、BDG、KL-6水平均较其治疗前下降,CD4^(+)/CD8^(+)水平均较其治疗前上升,且观察组患者治疗后的LDH、BDG及KL-6水平明显低于对照组,CD4^(+)/CD8^(+)水平明显高于对照组,差异均具有统计学意义(P<0.05);治疗后,两组患者的PaO_(2)、SaO_(2)水平均较其治疗前上升,且观察组患者治疗后的PaO_(2)水平明显高于对照组,差异均具有统计学意义(P<0.05),但两组患者治疗后的SaO_(2)水平比较差异无统计学意义(P>0.05);观察组患者的治疗总有效率为96.77%,明显高于对照组的74.19%,差异具有统计学意义(P<0.05);观察组和对照组患者的药物相关不良反应分别为9.68%、16.13%,差异无统计学意义(P>0.05)。结论复方磺胺甲恶唑联合卡泊芬净治疗可有效提高HIV感染合并PCP患者的临床效果,且安全性高。

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

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 (ALP) and type Ⅳ collagenase (matrix metalloproteinases, MMP 2, MMP 9) as well as the values of total protein (TP) and albumin (ALB) in BALF between the mild burden group of P carinii (involved alveoli <25% per 100 alveoli, Group 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 normal 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 <0 01); however, there were no differences in white cell differentiation Assays of biochemical markers showed that ALB in BALF in Group B (0 893±0 469?g/L) was increased in comparison with Group A (0 262±0 169 ?g/L); it was only 0 026±0 021?g/L in 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 and 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 <0 01) (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 burden of P carinii in lung tissues and the inflammatory reaction as well as biochemical markers of the resultant activity of lung injury

Iron chelator daphnetin against Pneumocystis carinii in vitro

Background Although there are several drugs and drug combinations for the treatment of Pneumocystis carinii (P. carinii) pne umonia, all drugs have the toxicity as well as low efficacy. Iron chelators have been proposed as a source of new drugs for combating these infections. We hypothesized that iron chelators would suppress the growth of P. carinii by deprivation of the nutritional iron required for growth. In this study, a short-term axenic culture system of P. carinii was established. Daphnetin (7,8-dihydroxycoumarin), a known iron chelator, was demonstrated to exhibit in vitro activity against P. carinii in this system. Methods P. carinii organisms were obtained from the lungs of immunosuppressed rats. The culture system consisted of Iscove Dulbecco Eagle’s Minimum Essential Medium (IMDM), supplemented with S-adenosyl-L-methionine, N-acetylglucosamine, putrescine, L-cysteine, L-glutamine, 2-mercaptoethanol, and fetal bovine serum, and was maintained at 37℃, in 5% CO 2, 95% O 2, at the optimal pH of 8.0. The culture system was used to assess the effect of daphnetin on the proliferation of P. carinii organisms. The ultrastructures of the treated organisms were observed by transmission electron microscopy.Results The number of cysts and trophozoites increased 8- to 9-fold and 11- to 12-fold, respectively, after 10 days of culture. Daphnetin was found to suppress the growth of P. carinii in a dose-dependent manner at concentrations between 1 μmol/L and 20 μmol/L. The inhibitory activity was suppressed by the chelation of daphnetin with ferrous sulfate in a 2∶1 molar ratio, but it was not suppressed by mixing the culture medium with magnesium sulfate. Reduction of P. carinii numbers after treatment with daphnetin correlated with morphological changes in the organisms, as determined by transmission electron microscopy.Conclusions Daphnetin can suppress the growth of P. carinii in vitro. The efficacy o f daphnetin in suppressing the the growth of P. carinii in vitro is related to its ability to chelate iron.

1例磺胺类药物过敏患者肝移植术后肺孢子菌感染的护理体会

本文总结1例磺胺类药物过敏患者肝移植术后肺孢子菌感染的护理经验。针对性进行磺胺脱敏治疗,从临床危险因素着手,开展病情观察、脱敏用药护理、过敏症状观察、正确雾化、咳嗽排痰、有效呼吸功能锻炼及康复训练、心理干预等护理干预,同时针对碳青霉烯类铜绿假单胞菌(CR-PA)做出相应的隔离措施,帮助患者树立治疗信心,促使积极配合治疗,加速疾病恢复。

非艾滋病肺孢子虫肺炎的临床特点分析

目的对非艾滋病肺孢子虫肺炎(PCP)患者的临床资料进行分析,总结其临床特点,从而提高临床诊治水平,改善患者预后。方法回顾性分析9例非艾滋病PCP患者的临床表现、实验室检查结果、影像学特征、治疗及转归。结果主要临床表现为发热7例(77.8%)、咳嗽6例(66.7%)、喘息4例(44.4%)。胸部CT主要表现为双肺弥漫性或散在斑片影、磨玻璃影及囊状透亮影,斑片影9例(100.0%)、磨玻璃影5例(55.6%)、囊状透亮影5例(55.6%),双肺内、中、外带分布无差异,肋膈角处可累及,合并纵隔淋巴结增大3例(33.3%)、胸腔积液1例(11.1%)。9例患者中,白细胞计数(WBC)升高5例(55.6%),平均WBC(10.53±4.24)×10^(9)/L;降钙素原(PCT)不同程度升高8例(88.9%),平均PCT(0.19±0.18)ng/ml;氧合指数不同程度下降7例(77.8%),平均氧合指数(245.50±140.30)mm Hg(1 mm Hg=0.133 kPa);淋巴细胞计数下降3例(33.3%),平均淋巴细胞计数(1.16±0.61)×10^(9)/L。6例(66.7%)应用复方磺胺甲噁唑(SMZ-TMP)+卡泊芬净治疗,1例(11.1%)单用SMZ-TMP,2例(22.2%)单用卡泊芬净,6例(66.7%)合并应用糖皮质激素。6例(66.7%)患者疗效好,3例(33.3%)患者病情恶化自动出院。结论非艾滋病PCP缺乏特异性临床特点,临床工作中需提高警惕,恶性肿瘤、长期接受激素治疗、糖尿病等免疫受损患者出现发热、咳嗽、喘息等临床症状,加之合并斑片影、磨玻璃影等胸部影像学改变,需及时留取合格下呼吸道标本送检,尽早干预治疗,改善预后。

AN EXPERIMENTAL TRIAL OF ARTEMETHER IN TREATMENT OF PNEUMOCYSTIS CARINII IN IMMUNOSUPPRESSED RATS

An immunosuppressed rat model was establisbed by injecting cortisone acetate 25 mg/rat twice a week for 4 weeks and 12.5mg/rat for another 2 weeks subcutaneously.A development of Pneumocystis carinii pneumonia(PCP) was found at the end of the 6th week in all rats.

白果内酯抗大鼠卡氏肺孢子虫肺炎的实验研究

目的 研究白果内酯治疗实验大鼠卡氏肺孢子虫肺炎的疗效。方法 以地塞米松皮下注射大鼠 6周诱导建立肺孢子虫肺炎动物模型 ,用不同剂量白果内酯治疗实验大鼠 ,以免疫抑制为对照组 ,通过大鼠肺质量 /体质量的值 ,肺印片每视野的包囊均数为指标评价疗效。结果 白果内酯 2 0mg/kg、3 0mg/kg治疗组大鼠肺质量 /体质量的值分别为 0 0 0 97± 0 0 0 3 3、0 0 10 2± 0 0 0 2 1,明显低于免疫抑制组对照组 0 0 14 6± 0 0 0 3 4(P <0 0 5 )。白果内酯 2 0mg/kg、3 0mg/kg治疗组大鼠肺印片每视野的包囊均数分别为 2 3 1± 0 88、1 95± 0 5 7明显低于免疫抑制组对照组 7 3 9± 2 3 3 (P <0 0 1)。

大蒜素治疗大鼠卡氏肺孢子虫肺炎的研究

目的 研究大蒜素对大鼠卡氏肺孢子虫肺炎 (PCP)的治疗效果。 方法 地塞米松连续肌肉注射Wistar大鼠 8周 ,建立大鼠PCP动物模型。用大蒜素治疗实验大鼠 ,同时设复方新诺明治疗对照组和PCP模型空白对照组。通过各组大鼠肺重、肺重 /体重比值、肺印片中每 10 0个油镜视野肺孢子虫包囊均数等指标考核疗效。 结果 大蒜素治疗组大鼠平均肺重为 1.73± 0 .17、肺重 /体重比值为 0 .84± 0 .12 ,显著低于PCP模型对照组的 2 .3 1± 0 .3 5、1.2 5± 0 .3 5 (P<0 .0 1)。肺印片中包虫数较PCP模型对照组减少 62 .9% ,其与复方新诺明治疗对照组接近。 结论 大蒜素对实验大鼠卡氏肺孢子虫肺炎有明显的治疗作用 ,治疗效果接近复方新诺明。

大鼠肺孢子虫肺炎动物模型的实验研究

目的建立肺孢子虫肺炎(PCP)的大鼠实验动物模型。方法将34只雌性清洁级SD大鼠随机分成实验组和对照组,每组17只。实验组采取每周2次每次每只皮下注射地塞米松1mg诱导的方法建立PCP动物模型;对照组则注射与地塞米松等体积的灭菌生理盐水。所有大鼠的饮水中加入1g/L盐酸四环素预防继发性细菌感染。12周后解剖全部大鼠,并分别制作肺组织印片,姬氏染色,检查肺孢子虫包囊。同时制作肺组织病理切片,HE染色,观察肺组织的病理学变化。结果用地塞米松诱导大鼠5周,肺组织印片均查见肺孢子虫包囊,肺组织也出现典型的病理学改变。连续诱导12周,均未见实验鼠死亡。实验组大鼠体重下降明显,与对照组比较,差异有非常显著性(P<0.01)。结论应用皮下注射地塞米松免疫抑制诱导的方法可成功建立PCP的大鼠动物模型。

半量诱导剂在建立卡氏肺孢子虫肺炎模型中的作用

目前，卡氏肺孢子虫肺炎（ＰＣＰ）模型的建立虽然可应用皮下注射可的松的方法获得稳定的效果，但是传统的诱导剂量存在着致实验动物死亡率高的问题。为此，本文采用减半诱导剂量的方法，既可使ＰＣＰ 模型建立，且又明显降低实验动物的死亡率，从而为ＰＣＰ的研究奠定较好的基础。

双氢青蒿素对卡氏肺孢子虫肺炎大鼠肺泡巨噬细胞凋亡的影响

目的 检测双氢青蒿素对卡氏肺孢子虫肺炎（PCP）大鼠肺泡巨噬细胞凋亡的影响。方法 以醋酸可的松皮下注射Wistar大鼠建立PCP动物模型,用60mg/kg双氢青蒿素治疗实验大鼠,杀鼠取肺,用胶原酶消化法分离肺泡巨噬细胞,用PI和TUNEL法检测其凋亡,同时设有正常大鼠对照组。结果 感染组和治疗组大鼠肺泡巨噬细胞凋亡率显著高于正常对照组,治疗组大鼠肺泡巨噬细胞凋亡率明显低于感染组。结论 卡氏肺孢子虫感染引起大鼠肺泡巨噬细胞发生凋亡,经双氢青蒿素治疗后PCP大鼠肺泡巨噬细胞凋亡降低。

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。