RESUMEN
Objective:To explore the correlation between protein energy wasting(PEW)and frailty in elderly patients on maintenance hemodialysis(MHD)and influencing factors of frailty.Methods:Clinical data of patients over 60 who had received regular hemodialysis treatment at Beijing Huairou Hospital between September to December 2021 were collected.According to Fried's evaluation criteria, patients were divided into a frailty group and a non-frailty group, and differences between the two groups were compared.Spearman correlation analysis was conducted to assess the correlation between protein energy wasting and frailty.Logistic regression was used to analyze the influencing factors of frailty in elderly patients on MHD.Results:A total of 81 MHD patients enrolled in this study, with 36 in the frailty group.The frailty group was older, had a higher proportion of patients with PEW, and underwent more months of dialysis, with a higher proportion of patients with diabetic nephropathy as the primary disease, having high levels of C-reactive protein(CRP), and having low KT/V, hemoglobin, albumin, body mass index(BMI), mid-arm circumference(MAC)and mid-arm muscle circumference(MAMC), and the differences with the control group were statistically significant(all P<0.05). The Fried frailty phenotype was positively correlated with age( r=0.021, P=0.047), but negatively associated with HGB( r=-0.329, P=0.003), albumin( r=0.021, P=0.047), BMI( r=0.021, P=0.047), TSF( r=-0.274, P=0.013), MAC( r=-0.554, P<0.001)and MAMC( r=-0.293, P=0.008). A Logistic regression equation was constructed using frailty as the dependent variable.The results showed that age, months of dialysis, KT/V, serum albumin and CRP were independent factors influencing the development of frailty in elderly patients with MHD. Conclusions:PEW and frailty coexist and interact with each other in elderly patients with MHD.Clinicians should place emphasis on the assessment of frailty and protein energy wasting in elderly dialysis patients and achieve early detection and intervention to avoid adverse clinical outcomes.
RESUMEN
Objective:To compare the clinicopathological differences between elderly and non-elderly patients with idiopathic membranous nephropathy(IMN).Methods:Patients diagnosed with IMN via renal biopsy at Beijing Huairou Hospital, Beijing Changping Hospital of Traditional Chinese Medicine, and Beijing Hospital of Traditional Chinese Medicine from January 2017 to August 2021 were retrospectively enrolled.They were classified into the elderly group(≥65 years)and the non-elderly group(<65 years), and the clinicopathological differences between the two groups were compared.Results:A total of 207 IMN patients were included in the study, with a male to female ratio of 1.7∶1.0.There were 56 patients in the elderly group, aged(68.2±3.1)years, and 151 patients in the non-elderly group, aged(48.2±6.2)years.Compared with the non-elderly group, the elderly group had a longer time from onset to renal biopsy and a higher proportion of patients with renal insufficiency and hypertension( P<0.05). The elderly group had a lower eGFR, lower serum albumin, higher serum cholesterol, and higher low-density lipoprotein than the non-elderly group( P<0.05). The proportions of patients with glomerulosclerosis, renal tubular atrophy, and interstitial fibrosis in the elderly group were higher than in the non-elderly group( P<0.05). The positive rates of glomerular PLA2R antigen staining in the two groups were 90.6%(29/32)and 91.0%(111/122), respectively, and there was no statistically significant difference between the two groups.IgG4 deposition represented the most common IgG subtype, with 93.8%(30/32)in the elderly group and 94.3%(115/122)in the non-elderly group.There was no statistical significance between the two groups( P>0.05). Conclusions:Compared with non-elderly IMN patients, a higher proportion of elderly IMN patients has renal insufficiency, hypertension and chronic renal pathology.The glomerular deposition of pathogenic antigens in elderly IMN patients was similar to that in non-elderly IMN patients, suggesting no difference in pathogenesis between the two groups.The clinicopathological differences between the two groups may be related to age and complications.
RESUMEN
Objective:To analyze characteristics and trends of histopathological diagnosis of adult renal biopsy in Beijing from 2008 to 2020.Methods:A total of 4 652 cases of adult renal biopsy were collected from three hospitals in Beijing between 2008 and 2020. The patients were divided into three age groups: 18-40 years, 40-65 years and≥ 65 years; and also divided into three period: 2008-2011, 2012-2015, and 2016-2020. The pathological characteristics and changes of renal biopsy were analyzed in three age groups at different periods.Results:Among 4 652 cases primary glomerular disease accounted for 81.8%, the membranous nephropathy (MN, 32.4%, 1 509/4 652), IgA nephropathy (IgAN, 29.2%, 1 356/4 652) and minor glomerular abnormalities (MGA, 11.3%, 526/4 652) were the top three pathological types. The overall proportion of MN and diabetic nephropathy (DN) increased from 20.3% and 2.3% in 2008-2011 to 37.3% and 10.1% in 2016-2020) (χ2=99.9 and 96.1, both P<0.01), respectively. For age group 18-40 years, the MN and DN increased from 11.2% and 1.6% in 2008-2011 to 24.7% and 5.5% in 2016-2020 (χ2=32.7 and 20.7, both P<0.01), respectively. For age group 40-65 years the MN and DN increased from 26.6% and 3.2% in 2008-2011 to 41.5% and 13.1% in 2016-2020 (χ2=39.1 and 57.3, both P<0.01), respectively. For age group≥65 years the MN was the most common pathological type in the three periods, fluctuating between 41.3% and 55.0% (χ2=5.2, P=0.08); and DN increased from 0(0/63) in 2008-2011 to 7.5%(22/292) in 2016-2020 (χ2=8.1, P=0.02). Conclusion:The renal biopsy data show that membranous nephropathy and diabetic nephropathy are the most common primary and secondary adult glomerular diseases in Beijing recently.
RESUMEN
OBJECTIVE:To explore clinical efficacy and safety of Ulinastatin injection combined with Xingnaojing injec-tion in the treament of severe craniocerebral injury(CCI). METHODS:A total of 120 severe CCI patients selected from our hospital during Sept. 2014-Nov. 2015 were divided into ulinastatin group,Xingnaojing group and combination group according to therapy plan,with 40 cases in each group. Three groups were given routine treatment timely after admission. On the basis of routine treatment,Ulinastatin group additionally received Ulinastatin injection 200 000 U,ivgtt,bid;Xingnaojing group addi-tionally received Xingnaojing injection 20 mL,ivgtt,qd;combination group additionally received Ulinastatin injection com-bined with Xingnaojing injection,same usage as above(with 1 h intervals). Three groups received therapy for consecutive 14 d. Serum inflammatory factors(CRP,IL-1,IL-6,TNF-α),serologic indexes of craniocerebral injury [neuron specific enolase (NSE),myelin basic protein(MBP),S100B protein(S100B)] and GCS scores before and after treatment as well as GOS scores after treatment were all observed in 3 groups. The occurrence of ADR was recorded during treatment. RESULTS:Before treatment,there was no statistical significance in serum inflammatory factors,serologic indexes of craniocerebral injury or GCS scores among 3 groups(P>0.05). Compared to before treatment,inflammatory factors of 3 groups were decreased signifi-cantly after treatment,the ulinastatin group was significantly lower than the Xingnaojing group,combination group was signifi-cantly lower than two single drug groups,with statistical significance(P<0.05). Levels of serologic indexes of craniocerebral injury and GCS scores of 3 groups were improved significantly,and the combination group was significantly better than the two single drug groups,with statistical significance(P<0.05). There was no statistical significance between ulinastatin group and Xingnaojing group(P>0.05). Six months after treatment,GOS score of combination group(4.17±0.81)was significantly better than those of ulinastatin group(3.05±0.97)and Xing-naojing group(2.97 ± 0.89),with statistical significance (P<0.05);there was no statistical significance between ulinastatin group and Xingnaojing group(P>0.05). During treatment,the incidence of ADR in combination group(27.50%)was significantly lower than ulinastatin group(50.00%)and Xingnaojing group(42.50%),with statistical significance(P<0.05);there was no statistical significance between ulinastatin group and Xingnaojing group(P>0.05). CONCLUSIONS:Ulinastatin injection combined with Xingnaojing injection can sig-nificantly decrease serum inflammatory factor levels,relieve craniocerebral injury,protect cerebral tissue and improve short-term prognosis with good safety.
RESUMEN
OBJECTIVE:To explore clinical efficacy and safety of Ulinastatin injection combined with Xingnaojing injec-tion in the treament of severe craniocerebral injury(CCI). METHODS:A total of 120 severe CCI patients selected from our hospital during Sept. 2014-Nov. 2015 were divided into ulinastatin group,Xingnaojing group and combination group according to therapy plan,with 40 cases in each group. Three groups were given routine treatment timely after admission. On the basis of routine treatment,Ulinastatin group additionally received Ulinastatin injection 200 000 U,ivgtt,bid;Xingnaojing group addi-tionally received Xingnaojing injection 20 mL,ivgtt,qd;combination group additionally received Ulinastatin injection com-bined with Xingnaojing injection,same usage as above(with 1 h intervals). Three groups received therapy for consecutive 14 d. Serum inflammatory factors(CRP,IL-1,IL-6,TNF-α),serologic indexes of craniocerebral injury [neuron specific enolase (NSE),myelin basic protein(MBP),S100B protein(S100B)] and GCS scores before and after treatment as well as GOS scores after treatment were all observed in 3 groups. The occurrence of ADR was recorded during treatment. RESULTS:Before treatment,there was no statistical significance in serum inflammatory factors,serologic indexes of craniocerebral injury or GCS scores among 3 groups(P>0.05). Compared to before treatment,inflammatory factors of 3 groups were decreased signifi-cantly after treatment,the ulinastatin group was significantly lower than the Xingnaojing group,combination group was signifi-cantly lower than two single drug groups,with statistical significance(P<0.05). Levels of serologic indexes of craniocerebral injury and GCS scores of 3 groups were improved significantly,and the combination group was significantly better than the two single drug groups,with statistical significance(P<0.05). There was no statistical significance between ulinastatin group and Xingnaojing group(P>0.05). Six months after treatment,GOS score of combination group(4.17±0.81)was significantly better than those of ulinastatin group(3.05±0.97)and Xing-naojing group(2.97 ± 0.89),with statistical significance (P<0.05);there was no statistical significance between ulinastatin group and Xingnaojing group(P>0.05). During treatment,the incidence of ADR in combination group(27.50%)was significantly lower than ulinastatin group(50.00%)and Xingnaojing group(42.50%),with statistical significance(P<0.05);there was no statistical significance between ulinastatin group and Xingnaojing group(P>0.05). CONCLUSIONS:Ulinastatin injection combined with Xingnaojing injection can sig-nificantly decrease serum inflammatory factor levels,relieve craniocerebral injury,protect cerebral tissue and improve short-term prognosis with good safety.
RESUMEN
Objective To investigate the effects of Paecilomyces Lilacinuson extracellular polysaccharides on the phenotypic and function maturity of mouse dendritic ceils.Methods Mononuclear cells were isolated from the mouse bone marrow cavity and added with cytokines for obtaining the recombinant mouse granulocyte-macrophagocyte colony stimulating factor(rmGM-CSF),recombinant mouse interleukin 4(rmIL-4) was induced to differentiated to immature DCs.Then different concentrations of extracellular polysaccharides were used to conduct the intervention.The mature DCs surface marker CD11c,major histocompatibility complex Ⅱ (MHC Ⅱ),CD80,CD86 molecular expression and phagocytosing FITC-dextran ability was detected by the flow cytometry.The effect of the polysaccharides on DCs Toll-like receptor(TLR)2 mRNA and TLR4 mRNA expression was detected by RT-PCR.Results After 400 μg/mL polysaccharides action for 48 h,the expression of DCs surface molecules such as CD11c,MHC Ⅱ,CD80 and CD86 was significantly up-regulated compared with the blank control group (P＜0.05);after the polysaccharides action,the ability of DCs phagocytosing FITC-dextran was decreased,especially the effects of 300,400 μg/mL of polysaccharides were more significant compared with the control group (P＜0.05).In addition,the polysaccharides could down-regulate the expression of TLR2 mRNA and TLR4 mRNA in DCs,the DCs down-regulation effect after 100-400 μg/mL polysaccharides treatment,the difference compared with the blank control group was statistically significant(P＜0.05).Conclusion The extracellular polysaccharides can up-regulate the expression of DCs surface CD11c,MHC Ⅱ,CD80 and CD 86 molecules,decreases the phagocytosis ability and down-regulates the expression of TLR2 mRNA and TLR4 mRNA,which preliminarily indicates that the polysaccharides could stimulate the differentiation and maturation of murine DCs.
RESUMEN
Objective:To investigate the effects of the Paecilomyces Lilacinuson extracellular polysaccharides on the phenotypic and maturation of murine dendritic cells. Methods: Imature DCs were induced in vitro from the murine bone marrow cells in the presence of rmGM-CSF and rmIL-4, and then they were cultured with different dosage of the extracellular polysaccharides. The morphological characterization was analyzed under microscopy. The expressions of the DCs surface costimulating factors and phagocytic function to FITC-dextran were detected by flow cytometry. The level of IL-12 secreted by DCs was observed by ELISA. At the same time the influence of DCs on the proliferation of T cells was determined by MTT. Results:Treating with the polysaccharides for 48 h could up-regulate the expression of DCs surface molecules,such as CD11c,MHCⅡ,CD80 and CD86,and the 400 μg/ml was the optimal dose,comparing with the blank control group, the difference was significant (P<0. 01), contrast to LPS control group that was not different ( P<0. 05 ) . The uptaking FITC-dextran ability of the DCs treated with 300 μg/ml and 400 μg/ml polysaccharides was significant lower than the unstimulated DCs(P<0. 05). At the same time the extract at different concentration could distinctly enhance the proliferation of T cells by DCs too. Conclusion:The extracellular polysaccharides could stimulate the maturation of dendritic cells and induce the production of mature dendritic cells.
RESUMEN
<p><b>OBJECTIVE</b>To observe the effects of warm needling moxibustion on body mass, knee cartilage andmorphology in rats with knee osteoarthritis (KOA).</p><p><b>METHODS</b>Forty SD rats were randomly divided into a normalgroup, a model group, a medication group and a warm needling group, 10 rats in each one. Except the normalgroup, the rats in the remaining three groups were injected with papain to establish the model of KOA. After themodeling, rats in the model group did not receive any treatment; rats in the warm needling group were treated withwarm needling moxibustion at bilateral "Xiqian"; rats in the medication group were treated with intragastric administration of meloxicam; rats in the normal group were treated with 0. 9% NaCl solution (identical dose as medication group) and immobilized as the warm needling group. The treatment was given once a day for consecutive20 days. The body mass, scale of knee cartilage and morphological changes were observed in each group after'treatment.</p><p><b>RESULTS</b>The increasing of body mass in the medication group and warm needling group was faster than!that in the model group, but slower than that in the normal group (all P<0. 05); the difference between medication group and warm needling group was not statistically significant (P>0. 05). The scale of knee cartilage in thewarm needling group and medication group was significantly lower than that in the model group (both P<0. 05),while the scale in the warm needling group was lower than that in the medication group (P<. 05). Regarding theknee morphology under micro-CT, the relief of knee degeneration and improvement of knee recovery in the warm needlinggroup were superior to those in the medication group.</p><p><b>CONCLUSION</b>The warm needling moxibustion could effectively reduce the knee pain, improve the recovery of knee cartilage, which is a safe and effective treatment.</p>
Asunto(s)
Animales , Humanos , Masculino , Ratas , Puntos de Acupuntura , Cartílago , Modelos Animales de Enfermedad , Articulación de la Rodilla , Moxibustión , Osteoartritis de la Rodilla , Terapéutica , Ratas Sprague-Dawley , Resultado del TratamientoRESUMEN
<p><b>OBJECTIVE</b>To compare the clinical efficacy on knee osteoarthritis (KOA) at the early and middle stage between electroacupuncture (EA) and meloxicam.</p><p><b>METHODS</b>Ninety patients of KOA at the early and middle stage were randomized into an EA group and a meloxicam group, 45 cases in each one. In the EA group, EA was applied to Dubi (ST 35), Neixiyan (EX-LE 4), Liangqiu (ST 34), Heding (EX-LE 2), Xuehai (SP 10), Yan- glingquan (GB 34) and Zusanli (ST 36); the needles were retained for 20 min and EA was applied once every two days. In the meloxicam group, the meloxicam tablets were prescribed for oral administration, 7. 5 mg, once a day. The treatment lasted for 6 weeks in the two groups. The Western Ontario and McMaster University Osteoarthritis Index (WOMAC) score, rectus femoris muscle tension, the 8-foot walking test and 5-time sit-to-stand test were adopted to observe and compare the effects in the two groups.</p><p><b>RESULTS</b>After treatment, every item score in WOMAC was reduced after treatment (all P < 0.05), but the difference was not significant between the two groups (all P > 0.05). In the EA group, the rectus femoris tension after treatment was reduced as compared with that before treatment (P < 0.05) and the reducing result was much more apparent as compared with that in the meloxicam group (P < 0.05). For the 8-foot walking test and 5-time sit-to-stand test, the time was shortened after treatment in the two groups (all P < 0.05) and the result in the EA group was much more obvious than that in the meloxicam group (both P < 0.05).</p><p><b>CONCLUSION</b>Both EA and meloxicam are effective in the treatment of KOA at the early and middle stage. EA improves rectus femoris tension and recovers the internal mechanics balance and the efficacy is better than that of meloxicam.</p>
Asunto(s)
Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Puntos de Acupuntura , Electroacupuntura , Osteoartritis de la Rodilla , Quimioterapia , Terapéutica , Tiazinas , Usos Terapéuticos , Tiazoles , Usos Terapéuticos , Resultado del TratamientoRESUMEN
BACKGROUND:Studies have shown that signaling pathway plays an important role in the pathogenesis of knee osteoarthritis. OBJECTIVE: To elaborate the experimental progress of signal transduction pathway in knee osteoarthritis METHODS: A computer-based retrieval of PubMed, CNKI, VIP and Wanfang databases was performed to find the relevant literatures concerning experimental studies of signaling pathway in knee osteoarthritis. Al data were primarily screened to exclude repeated and irrelevant articles. Articles about association between knee osteoarthritis and signal pathway were included.RESULTS AND CONCLUSION:A total of 51 articles were colected, including 20 in Chinese and 31 in English. It is concluded that the signal transduction pathway related to knee osteoarthritis is mainly concentrated in the two aspects: the proliferation of chondrocytes and the apoptosis of chondrocytes. The signal transduction pathways related to the proliferation of chondrocytes mainly include Notch signaling pathway and Wnt signaling pathway. SDF-1/CXCR4 signaling pathway, TLR4 signaling pathway, and MAPKs signaling pathway are associated with apoptosis in chondrocytes. Hippo-YAP signaling pathway and ERK1/2 signaling pathway have a double role in the regulation of proliferation and apoptosis of chondrocytes.
RESUMEN
BACKGROUND:Studies have shown that the non-surgical therapy for knee osteoarthritis is exactly effective, but there is a lack of scientific and rational evaluation system. OBJECTIVE:To review the evaluation methods of non-surgical therapy for knee osteoarthritis in recent 5 years. METHODS:A computer-based retrieval of PubMed, WOK, CNKI, VIP, Wanfang databases was performed to find literature related to the evaluation methods of non-surgical therapy for knee osteoarthritis. Al data were primarily screened to exclude irrelevant literature. Those literatures about the evaluation methods of non-surgical therapy for knee osteoarthritis were included. Repetitive studies and untypical reports were excluded. RESULTS AND CONCLUSION:Total y 42 articles were col ected, including 28 in Chinese and 14 in English. The analysis results showed that the non-surgical treatment for knee osteoarthritis can improve knee function and the quality of life in patients, and have an exact effect and a good economic benefit. Therefore, exploring a scientific and reasonable evaluation method to guide the choice of clinical treatment for knee osteoarthritis is greatly significant, which can improve the efficacy of non-surgical therapy for knee osteoarthritis. Scale-based evaluation method is simple and practical, but the presence of a single scale has a lack of objectivity. The method of biomechanics or imaging has the advantages of objective, highly reliable, accurate, non-invasive and so on. In the future therapeutic evaluation system, the combination of subjective scale observation and objective indicators should be more recommended, and the evaluation methods of non-surgical therapy for knee osteoarthritis should be selected appropriately based on the difference of clinical therapy and effect.
RESUMEN
Objective To explore the relationship of body mass index ( BMI ) and atherosclerosis,blood pressure( BP) in patients with maintenance hemodialysis(MHD),to further elucidate the pathogenesis of cardiovascular disease in MHD patients.Methods The relationship between BP,intima-medial thickness (IMT) and BMI was analyzed in 101 patients with MHD.Results After dialysis,38％ patients had systolic blood pressure over 150mm Hg,21％ had diastolic blood pressure of more than 90mm Hg.Contrary to the general population,the correlation between BP and BMI were negatively (r =-0.13,P ＜ 0.01 ).Hypertension in patients with BMI ＜ 20 were more than that in patients with a normal body weight ( BMI 20.0 ～ 27.5 ) and overweight patients( BML ＞ 27.5 ),and the rate was 35％,29％ and 20％,respectively.Univariate analysis showed that BP and BMI were negatively correlated(r =-0.12,P ＜0.01),IMT was positively correlated with BMI(r=0.411,P ＜0.05).Conclusion In persistent hemodialysis patients,malnutrition was correlated with the occurrence and development of atherosclerosis and hypertension,and BMI decreasing was an independent risk factor,which could increase the morbidity and mortslity of cardiovascular disease.
RESUMEN
Malignant gliomas are the most common intracranial tumors,with a highly invasive nature and a poor prognosis. The invasion of malignant gliomas is a complicate and consecutive multi-step process that includes adhesion, degradation and migration and involves various factors such as cell adhesion molecules, enzymes, and cytokines. Research on the invasive properties of malignant gliomas will advance our understanding of their underlying biological behavior and improve clinical treatment.
RESUMEN
Objective:To investigate the quantification of CD4~+CD25~+ regulatory T cells and distribution of CD4~+CD8~+ T lymphocyte subgroup in peripheral blood of patients in chronic hepatitis B (CHB),and to reveal relationship between CD4~+CD25~+ regulatory T cells,CD4~+CD8~+ T lymphocyte subgroup and HBV infetion as well.Methods:CD4~+CD25~(high),CD4~+CD25~+Foxp3~+Treg and CD3~+CD4~+CD8~+T lymphocyte subgroup in peripheral blood from 50 patients with CHB and 20 healthy controls was analyzed using flow cytometry.HBV DNA was detected by fluorescence quantitative PCR.Results:The number of CD4~+CD25~(high)Tregs in patients with CHB was obviously higher than that in healthy controls(P＜0.01)and increased with copies of HBV DNA.The same with the change of CD4~+CD25~+Foxp3~+Tregs in patients with CHB and there was a positive correlation between CD4~+CD25~(high)Tregs and CD4~+CD25~+Foxp3~+Tregs(r=0.890,P＜0.001).Compared with healthy controls,the frequency of CD4~+T cells and the ratio of CD4~+/CD8~+ in patients with CHB was declined,but there was no significant difference in the frequency of CD3~+T cells and CD8~+T cells between them(P＞0.05).The variation in the number of CD4~+CD25~(high)Tregs was correlated positively with the copies of HBV DNA(r=0.782,P＜0.001)and glutamic-pyruvic transaminase(ALT)(r=0.432,P＜0.005)separately,but negatively with the frequency of CD3~+,CD4~+,CD8~+T cells and the ratio of CD4~+/CD8~+(P＞0.05).The variation in the frequency of CD3~+,CD4~+,CD8~+T cells and the ratio of CD4~+/CD8~+ was also correlated negatively with the copies of HBV DNA(P＞0.05).Conclusion:The number of CD4~+CD25~(high)Tregs increases in patients with CHB and is in accordance with the copies of HBV DNA and increased level of ALT.Further studies should be done to investigate weather CD4~+CD8~+ T lymphocyte subgroup could be used to monitor the state of community.
RESUMEN
Objective:To construct the plant expression vector of Der f1 allergen of Dermatophagoides pteronyssinu and expression in tobacco lamina.Methods:The Der f1 gene was amplified from the glycerin bacterium which contained pET28a(+)-Der f1 plasmid,cloned into the pMD 19-T plasmid,and then sequenced.The Der f1 gene was digested by ClaⅠand SalⅠ,and cloned into potato virus X (PVX) to construct plant expression vector PVX-Der f1,and then was transformed agrobacterium tumefaciens.The positive one was selected to infect tobacco lamina for expressing target protein.The protein was identified and analysed by SDS-PAGEand Western blot.Results:Digestion and sequence analysis confirmed that the plant expression vector was correct,and the SDS-PAGE and Western blot results showed that the molecular weight of the protein was about 34M_r and it could specific binding with positive serum.Conclusion:The plant expression vector of Der f1 is successfully constructed and the recombinant protein is also produced.
RESUMEN
Objective To detect the expression level of stage-specific genes in Leishmania promastigotes and amastigotes. Methods Total RNAs were isolated from Leishmania amazonensis stationary promastigotes and three sources of amastigotes: freshly obtained from mouse skin lesions,infected J774.G8 macrophages,and transformed from the cultured promastigotes. mRNAs were conversely transcribed into cDNA with SuperScripII reverse transcriptase and oligo dT primers. The polymerase chain reaction(PCR)was used to amplify the specific fragments of amastigote-specific nuclease(P-4)and promastigote-specific membrane glycoprotein(GP-46). PCR products were analyzed in 1.5% agarose gel. Results A P-4-specific band(273 bp)was observed in all three types of amastigotes with similar density,but it was almost undetectable in promastigotes. In contract,a GP-46-specific band(325 bp)was expressed at a higher level in promastigotes than in all three types of amastigotes. Conclusion Promastigote-derived amastigotes express high level of P-4-specific gene and can be used as a source of amastigotes for biochemical and immunological studies.