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(p55-58...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.展开更多
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% so...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.展开更多
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 ...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.展开更多
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 Ri...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.展开更多
Objective To understand the interaction between surfactant proteins and pneumocystis carinii pneumonia (PCP),and the impact of corticosteriods on surfactant proteins.Methods We established rat models of PCP and bacter...Objective To understand the interaction between surfactant proteins and pneumocystis 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 fluid(BALF)of rats was collected.Total nucleated cells of BALF were counted and differentiated,and the concentrations of surfactant protein A(SP-A)and surfactant 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 control(n = 6),consisted of rats with cortisone acetate injection for over 8 wk without lung infection;group Ⅲ,bacterial pneumonia(n = 11),rats were injected with cortisone acetate over 8 wk that resulted in bacterial pneumonia without other pathogens isolated;and group Ⅳ,PCP(n = 14),rats with injected cortisone acetate 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 ＜ 0.001).During PCP infection,the total cell count and the percentage of polymorphonuclearcytes(PMNs)in BALF were significantly increased(P ＜ 0.01),but were lower than those in the bacterial pneumonia 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 ＜ 0.05)and bacterial pneumonia groups(6.2 ± 5.6 μg/ml,P ＜ 0.001).We also found that the relative content of SP-D was significantly higher in PCP(24249 ±4780 grey values)than that in the negative control (13 384 ± 2887 grey values,P ＜ 0.001)and that in bacterial pneumonia(11 989 ± 2750 grey values,P＜0.001).SP-A and SP-D were also higher in the moderate to heavy group of PCP than those seen in the mild group(P ＜ 0.01,P ＜ 0.001).SP-A and SP-D were higher in the negative control group than those in the normal control group,but there was no significant difference between the 2 groups.Conclusion These results suggest that the concentrations of SP-A and SP-D in BALF are increased by pneumocystis carinii specific stimulation,but the alteration is not related to the corticosteriod usage.展开更多
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 pn...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展开更多
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 prop...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.展开更多
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) wa...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.展开更多
基金supported by a grant from a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘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.
文摘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.
文摘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.
文摘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.
基金ThisresearchwassupportedbygrantsfromThetrainingprojectoftheShanghaiHealthSystem (No 98BR0 3 0 )andtheShanghaiEducationCommittee (No 98QN2 7)
文摘Objective To understand the interaction between surfactant proteins and pneumocystis 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 fluid(BALF)of rats was collected.Total nucleated cells of BALF were counted and differentiated,and the concentrations of surfactant protein A(SP-A)and surfactant 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 control(n = 6),consisted of rats with cortisone acetate injection for over 8 wk without lung infection;group Ⅲ,bacterial pneumonia(n = 11),rats were injected with cortisone acetate over 8 wk that resulted in bacterial pneumonia without other pathogens isolated;and group Ⅳ,PCP(n = 14),rats with injected cortisone acetate 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 ＜ 0.001).During PCP infection,the total cell count and the percentage of polymorphonuclearcytes(PMNs)in BALF were significantly increased(P ＜ 0.01),but were lower than those in the bacterial pneumonia 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 ＜ 0.05)and bacterial pneumonia groups(6.2 ± 5.6 μg/ml,P ＜ 0.001).We also found that the relative content of SP-D was significantly higher in PCP(24249 ±4780 grey values)than that in the negative control (13 384 ± 2887 grey values,P ＜ 0.001)and that in bacterial pneumonia(11 989 ± 2750 grey values,P＜0.001).SP-A and SP-D were also higher in the moderate to heavy group of PCP than those seen in the mild group(P ＜ 0.01,P ＜ 0.001).SP-A and SP-D were higher in the negative control group than those in the normal control group,but there was no significant difference between the 2 groups.Conclusion These results suggest that the concentrations of SP-A and SP-D in BALF are increased by pneumocystis carinii specific stimulation,but the alteration is not related to the corticosteriod usage.
基金ThisworkwassupportedbytheTrainingProjectofShanghaiHealthSystem (No 98BR0 30 )andShanghaiEducationCommittee(No 98QN2 7)
文摘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
文摘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.
文摘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.
