Objective. So far, it has not been established a satisfactory method for early diagnosis and studying on epidemiology for leprosy, we want to develop a molecular biological method for solv...Objective. So far, it has not been established a satisfactory method for early diagnosis and studying on epidemiology for leprosy, we want to develop a molecular biological method for solving this point. Materials and methods. Based on the M. leprae gene coding groEL, 65kD and 16S rRNA, three polymerase chain reactions were developed by using plikaytis’, Woods’ and Pattyn’s procedures. It was optimized that the experimental parameters for each PCR, and a comparative study on practivity among three PCRs was also conducted for practical purpose.Results and conclusion. For detecting infection with M. leprae, all of PCRs established by us were highly sensitive and specific, but for practicl purpose, the woods’ PCR optimized by us ought to be chosen firstly.展开更多
Immune-deficient nude mice were inoculated with nude mouse derived Mycobacteriurn leprae by multiple routes (intravenously. subcutaneously at the foot pads and ears). The results showed that these inoculated animals w...Immune-deficient nude mice were inoculated with nude mouse derived Mycobacteriurn leprae by multiple routes (intravenously. subcutaneously at the foot pads and ears). The results showed that these inoculated animals were capable of producing a great number of Mycobacterium leprae to a level 10(11-12) per gram of tissue. and were detected histopathologically to have heavy leproniatous lesions. The dissemination of the infection was found particularly in sites with lower body temperature.The organisms have a partiality to striated muscles and peripheral nerves. The authors suggest that experimental leprosy in nude mice is a very useful tool for leprosy resarch. especially in countries without armadillos. Compared with the single-route inoculation reported previously. multiple-route inoculation is more available.展开更多
INTRODUCTIONLeprosy caused by Mvcobacterium leprae （M. leprae）, is a chronic granulomatous disease affecting the skin and peripheral nervous system, which is transmitted through direct contact with nontreated or ina...INTRODUCTIONLeprosy caused by Mvcobacterium leprae （M. leprae）, is a chronic granulomatous disease affecting the skin and peripheral nervous system, which is transmitted through direct contact with nontreated or inadequate treatment patients. Diagnosis of leprosy depends on the clinical signs and symptoms and slit skin smear positivity. However, it＇s sometimes similar with other granulomatous disease caused by mycobacterial infection. Early stage leprosy is difficult to diagnose by clinical criterion alone because the sensitivity of acid-fast bacilli staining is quite low. The polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） shows the great advantage in rapid identification and diagnosis for early cases and has a differentiation between leprosy and nonleprosy cases.展开更多
Objective:This study was performed to compare the efficacy and safety of methotrexate(MTX)plus prednisolone versus prednisolone monotherapy in patients with erythema nodosum leprosum(ENL).Methods:This comparative clin...Objective:This study was performed to compare the efficacy and safety of methotrexate(MTX)plus prednisolone versus prednisolone monotherapy in patients with erythema nodosum leprosum(ENL).Methods:This comparative clinical study was performed in the Chittagong Medical College Hospital,Bangladesh from June 2018 to December 2019.Nineteen patients were selected according to inclusion criteria and randomly allocated to either the MTX plus prednisolone group(Group A,n=10)or the prednisolone monotherapy group(Group B,n=9).All patients were followed up until the end of the 6-month duration of therapy to observe the clinical outcomes and adverse effects.Results:All patients in both groups showed significant improvement(P=0.005 and P=0.008 in Group A and B,respectively).However,prednisolone-related adverse events were more frequent in Group B.Conclusion:The present study has demonstrated that a combination of prednisolone and MTX is a safer and more effective treatment than steroid monotherapy in ENL patients including the healing of nodules.However,this combination therapy did not appear to have a significant steroid-sparing effect,possibly because of the small sample size and short study period.Therefore,a well-designed multicenter randomized controlled trial is recommended for validation of MTX with prednisolone for the management of ENL.展开更多
BACKGROUND Leprosy is a disease caused by Mycobacterium leprae(M.leprae),an intracellular pathogen that has tropism and affects skin and nervous system cells.The disease has two forms of presentation:Paucibacillary an...BACKGROUND Leprosy is a disease caused by Mycobacterium leprae(M.leprae),an intracellular pathogen that has tropism and affects skin and nervous system cells.The disease has two forms of presentation:Paucibacillary and multibacillary,with different clinical and immunological manifestations.Unlike what occurs in the multibacillary form,the diagnostic tests for the paucibacillary form are nonspecific and not very sensitive,allowing the existence of infected individuals without treatment,which contributes to the spread of the pathogen in the population.To mitigate this contamination,more sensitive diagnostic tests capable of detecting paucibacillary patients are needed.AIM To predict the three-dimensional structure models of M.leprae antigens with serodiagnostic potential for leprosy.METHODS In this in silico study,satisfactory templates were selected in the Protein Data Bank(PDB)using Basic Local Alignment Search Tool to predict the structural templates of ML2038,ML0286,ML0050,and 85B antigens by comparative modeling.The templates were selected according to general criteria such as sequence identity,coverage,X-ray resolution,Global Model Quality Estimate value and phylogenetic relationship;Clustal X 2.1 software was used in this analysis.Molecular modeling was completed using the software Modeller 9v13.Visualization of the models was made using ViewerLite 4.2 and PyMol software,and analysis of the quality of the predicted models was performed using the QMEAN score and Z-score.Finally,the three-dimensional moels were validated using the MolProbity and Verify 3D platforms.RESULTS The three-dimensional structure models of ML2038,ML0286,ML0050,and 85B antigens of M.leprae were predicted using the templates PDB:3UOI(90.51%identity),PDB:3EKL(87.46%identity),PDB:3FAV(40.00%identity),and PDB:1F0N(85.21%identity),respectively.The QMEAN and Z-score values indicated the good quality of the structure models.These data refer to the monomeric units of antigens,since some of these antigens have quaternary structure.The validation of the models was performed with the final three-dimensional structure-monomer(ML0050 and 85B antigens)and quaternary structures(ML2038 and ML0286).The majority of amino acid residues were observed in favorable and allowed regions in the Ramachandran plot,indicating correct positioning of the side chain and absence of steric impediment.The MolProbity score value and Verify 3D results of all models indicated a satisfactory prediction.CONCLUSION The polarized immune response against M.leprae creates a problem in leprosy detection.The selection of immunodominant epitopes is essential for the development of more sensitive serodiagnostic tests,for this it is important to know the three-dimensional structure of the antigens,which can be predicted with bioinformatics tools.展开更多
目的:了解云南、四川、贵州三省麻风菌株基因型的地理分布。方法:从云南省丘北县、四川省凉山州、贵州省兴义市分别采集59、21、31例麻风患者皮损活检,提取麻风菌DNA,对6-7、AC9、GTA9 3个数目可变的串联重复(variable-number tandem re...目的:了解云南、四川、贵州三省麻风菌株基因型的地理分布。方法:从云南省丘北县、四川省凉山州、贵州省兴义市分别采集59、21、31例麻风患者皮损活检,提取麻风菌DNA,对6-7、AC9、GTA9 3个数目可变的串联重复(variable-number tandem repeat,VNTR)位点的PCR产物测序,确定各位点重复序列数进行分型。结果:三省在6-7位点上有5种基因型,云南以7个拷贝的基因型为主,其它两地区的菌株在该位点的等位基因型为7、8、9,其分布无显著性差异。AC9位点上有4种基因型,三省均以8个拷贝的基因型为主。GTA9位点有非常明显的多样性。云南GTA9为9个拷贝的菌株占总菌株数的18.18%。结论:本研究结果提示麻风菌的基因型与地理分布有关。但需要以更多的位点,了解以VNTR为主的基因分型能否应用于麻风病的传染源和传播链的流行病学研究。展开更多
文摘Objective. So far, it has not been established a satisfactory method for early diagnosis and studying on epidemiology for leprosy, we want to develop a molecular biological method for solving this point. Materials and methods. Based on the M. leprae gene coding groEL, 65kD and 16S rRNA, three polymerase chain reactions were developed by using plikaytis’, Woods’ and Pattyn’s procedures. It was optimized that the experimental parameters for each PCR, and a comparative study on practivity among three PCRs was also conducted for practical purpose.Results and conclusion. For detecting infection with M. leprae, all of PCRs established by us were highly sensitive and specific, but for practicl purpose, the woods’ PCR optimized by us ought to be chosen firstly.
文摘Immune-deficient nude mice were inoculated with nude mouse derived Mycobacteriurn leprae by multiple routes (intravenously. subcutaneously at the foot pads and ears). The results showed that these inoculated animals were capable of producing a great number of Mycobacterium leprae to a level 10(11-12) per gram of tissue. and were detected histopathologically to have heavy leproniatous lesions. The dissemination of the infection was found particularly in sites with lower body temperature.The organisms have a partiality to striated muscles and peripheral nerves. The authors suggest that experimental leprosy in nude mice is a very useful tool for leprosy resarch. especially in countries without armadillos. Compared with the single-route inoculation reported previously. multiple-route inoculation is more available.
基金The study was supported by a grant from the National Natural Science Foundation of China （No. 81271761）.
文摘INTRODUCTIONLeprosy caused by Mvcobacterium leprae （M. leprae）, is a chronic granulomatous disease affecting the skin and peripheral nervous system, which is transmitted through direct contact with nontreated or inadequate treatment patients. Diagnosis of leprosy depends on the clinical signs and symptoms and slit skin smear positivity. However, it＇s sometimes similar with other granulomatous disease caused by mycobacterial infection. Early stage leprosy is difficult to diagnose by clinical criterion alone because the sensitivity of acid-fast bacilli staining is quite low. The polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） shows the great advantage in rapid identification and diagnosis for early cases and has a differentiation between leprosy and nonleprosy cases.
基金The authors thankfully acknowledge the assistance of Mr. Sultan Md Elias Uddin. Program Manager, Leprosy control project, Chittagong.
文摘Objective:This study was performed to compare the efficacy and safety of methotrexate(MTX)plus prednisolone versus prednisolone monotherapy in patients with erythema nodosum leprosum(ENL).Methods:This comparative clinical study was performed in the Chittagong Medical College Hospital,Bangladesh from June 2018 to December 2019.Nineteen patients were selected according to inclusion criteria and randomly allocated to either the MTX plus prednisolone group(Group A,n=10)or the prednisolone monotherapy group(Group B,n=9).All patients were followed up until the end of the 6-month duration of therapy to observe the clinical outcomes and adverse effects.Results:All patients in both groups showed significant improvement(P=0.005 and P=0.008 in Group A and B,respectively).However,prednisolone-related adverse events were more frequent in Group B.Conclusion:The present study has demonstrated that a combination of prednisolone and MTX is a safer and more effective treatment than steroid monotherapy in ENL patients including the healing of nodules.However,this combination therapy did not appear to have a significant steroid-sparing effect,possibly because of the small sample size and short study period.Therefore,a well-designed multicenter randomized controlled trial is recommended for validation of MTX with prednisolone for the management of ENL.
文摘BACKGROUND Leprosy is a disease caused by Mycobacterium leprae(M.leprae),an intracellular pathogen that has tropism and affects skin and nervous system cells.The disease has two forms of presentation:Paucibacillary and multibacillary,with different clinical and immunological manifestations.Unlike what occurs in the multibacillary form,the diagnostic tests for the paucibacillary form are nonspecific and not very sensitive,allowing the existence of infected individuals without treatment,which contributes to the spread of the pathogen in the population.To mitigate this contamination,more sensitive diagnostic tests capable of detecting paucibacillary patients are needed.AIM To predict the three-dimensional structure models of M.leprae antigens with serodiagnostic potential for leprosy.METHODS In this in silico study,satisfactory templates were selected in the Protein Data Bank(PDB)using Basic Local Alignment Search Tool to predict the structural templates of ML2038,ML0286,ML0050,and 85B antigens by comparative modeling.The templates were selected according to general criteria such as sequence identity,coverage,X-ray resolution,Global Model Quality Estimate value and phylogenetic relationship;Clustal X 2.1 software was used in this analysis.Molecular modeling was completed using the software Modeller 9v13.Visualization of the models was made using ViewerLite 4.2 and PyMol software,and analysis of the quality of the predicted models was performed using the QMEAN score and Z-score.Finally,the three-dimensional moels were validated using the MolProbity and Verify 3D platforms.RESULTS The three-dimensional structure models of ML2038,ML0286,ML0050,and 85B antigens of M.leprae were predicted using the templates PDB:3UOI(90.51%identity),PDB:3EKL(87.46%identity),PDB:3FAV(40.00%identity),and PDB:1F0N(85.21%identity),respectively.The QMEAN and Z-score values indicated the good quality of the structure models.These data refer to the monomeric units of antigens,since some of these antigens have quaternary structure.The validation of the models was performed with the final three-dimensional structure-monomer(ML0050 and 85B antigens)and quaternary structures(ML2038 and ML0286).The majority of amino acid residues were observed in favorable and allowed regions in the Ramachandran plot,indicating correct positioning of the side chain and absence of steric impediment.The MolProbity score value and Verify 3D results of all models indicated a satisfactory prediction.CONCLUSION The polarized immune response against M.leprae creates a problem in leprosy detection.The selection of immunodominant epitopes is essential for the development of more sensitive serodiagnostic tests,for this it is important to know the three-dimensional structure of the antigens,which can be predicted with bioinformatics tools.
文摘目的:了解云南、四川、贵州三省麻风菌株基因型的地理分布。方法:从云南省丘北县、四川省凉山州、贵州省兴义市分别采集59、21、31例麻风患者皮损活检,提取麻风菌DNA,对6-7、AC9、GTA9 3个数目可变的串联重复(variable-number tandem repeat,VNTR)位点的PCR产物测序,确定各位点重复序列数进行分型。结果:三省在6-7位点上有5种基因型,云南以7个拷贝的基因型为主,其它两地区的菌株在该位点的等位基因型为7、8、9,其分布无显著性差异。AC9位点上有4种基因型,三省均以8个拷贝的基因型为主。GTA9位点有非常明显的多样性。云南GTA9为9个拷贝的菌株占总菌株数的18.18%。结论:本研究结果提示麻风菌的基因型与地理分布有关。但需要以更多的位点,了解以VNTR为主的基因分型能否应用于麻风病的传染源和传播链的流行病学研究。
