In recent years, valuable experience and insights have been gained into L-forms (cell-wall-deficient variants) of bacteria and fungi and their disease-trigger potential in cases with chronic infections, autism spectru...In recent years, valuable experience and insights have been gained into L-forms (cell-wall-deficient variants) of bacteria and fungi and their disease-trigger potential in cases with chronic infections, autism spectrum disorders, autoimmune and neurodegenerative diseases. Based on the concept of “internal” blood microbiota, consisting of L-forms and its relevance to health and disease, the current study aims to outline the profile of dysbiotic disorders in three cancer patients (with endometrial cancer, breast cancer and acute myeloid leukemia), all in a phase before chemotherapy. Venous blood samples from the patients and from one control healthy person, were microbiologically studied. The used novel methodology of blood microbiota assessment was based on the following phases: isolation of L-forms, development and propagation, cultivation and conversion of L-forms into classical bacteria and fungi, as well as their identification with MALDI-TOF method. From the patients were isolated L-forms of opportunistic bacteria (Enterococcus faecalis, Esherichia coli, Enterobacter cloacae and Pseudomonas oryzihabitans) and fungi such as Rhodotorula mucilaginosa, Aspergillus fumigatus and Mucorales. In conclusion, the common feature found for the three cancer patients was the isolation from the blood of highly associated communities consisting of morphologically indistinguishable L-bodies, which through reversion in broth, were identified as distinct bacterial and fungal species. Unlike classic bacteria or fungi causing sepsis and bacteremia/fungemia, the presence of L-forms in blood is hidden, it does not demonstrate clinical signs nor it can be detected by conventional methods. It should be noted, however, that the dysbiotic blood microbiota shows unique and individual characteristics for the concrete cancer patient, correlates to the common state of the organism and tumor localization in the body, as well as it outlines the cancer promoting role of L-forms in processes of malignization, cancer genesis and progression.展开更多
<b><span style="font-family:Verdana;">Background</span></b><span style="font-family:""><span style="font-family:Verdana;">: The recognition of hum...<b><span style="font-family:Verdana;">Background</span></b><span style="font-family:""><span style="font-family:Verdana;">: The recognition of human blood microbiota, consisting of cell wall-deficient microbes (L-forms), is a major challenge today in the field of microbiology. There are accumulating data confirming the concept of “internal” blood L-form microbiota and its significance for health and diseases. Finding out whether the blood microbiota can be of diagnostic and prognostic importance for detection and evaluation of chronic infections anywhere in </span><span style="font-family:Verdana;">the body is a major objective. In the context of chronically infected upper</span> <span style="font-family:Verdana;">respiratory tract (URT), the aim of the current study was to understand</span><span style="font-family:Verdana;"> wheth</span><span style="font-family:Verdana;">er a local infection can be a source for entry of bacteria and fungi in th</span><span style="font-family:Verdana;">e blood. </span><b><span style="font-family:Verdana;">Methods: </span></b><span style="font-family:Verdana;">Blood samples from six persons with chronic inflammations</span><span style="font-family:Verdana;"> in URT diagnosed with hypertrophied adenoids, chronic sinusitis, nasal polyps, chronic naso-pharyngitis and one control healthy person were studied. Blood microbiota assessment methodology that be used, included three phases: </span></span><span style="font-family:Verdana;">1</span><span style="font-family:""><span style="font-family:Verdana;">) </span><span style="font-family:Verdana;">isolation of L-form cultures from blood-development and propagation;</span></span><span style="font-family:Verdana;">2</span><span style="font-family:""><span style="font-family:Verdana;">) cultivation directed to conversion of L-forms into bacterial and fungal cul</span><span style="font-family:Verdana;">tures;</span></span><span style="font-family:Verdana;">3</span><span style="font-family:Verdana;">) isolation of pure classical bacterial and fungal cultures and their</span><span style="font-family:""> <span><span style="font-family:Verdana;">identification by MALDI-TOF method. </span><b><span style="font-family:Verdana;">Results:</span></b><span style="font-family:Verdana;"> From the patients were isolated </span></span><span style="font-family:Verdana;">L-forms of opportunistic bacteria (</span><i><span style="font-family:Verdana;">Streptococcus mitis</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Roseomonas mucosa</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Dermacoccus nishinomiyaensis</span></i><span><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Enterococcus faecalis</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Acinetobacter johnsonii</span></i><span style="font-family:Verdana;">, </span></span><i><span style="font-family:Verdana;">Pseudomonas putida</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Staphylococcus aureus</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Pseudomonas luteola</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Enterobacter cloacae</span></i><span style="font-family:Verdana;">) and fungi such as </span><i><span style="font-family:Verdana;">Rhodotorula mucilaginosa</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Aspergillus niger</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Aspergillus fumigatus and Mucorales.</span></i> <b><span style="font-family:Verdana;">Conclusion:</span></b><span style="font-family:Verdana;"> The novel innovative methodology for assessment of blood L-form microbiota was successfully applied for detection of microbes responsible for chronic infections at URT.展开更多
Objective: To study the relationship between the polymorphism of drug resistant gene rpoB and drug resistance against rifampicin(RFP) of M. tuberculosis L-forms, and to evaluate its clinical application. Methods: A to...Objective: To study the relationship between the polymorphism of drug resistant gene rpoB and drug resistance against rifampicin(RFP) of M. tuberculosis L-forms, and to evaluate its clinical application. Methods: A total of 52 clinical isolated strains of M. tuberculosis L-forms were collected. rpoB gene polymorphism was analyzed by polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP) and conventional antimicrobial susceptibility test (AST). Their results were compared. Results: AST results showed that 38 of 52 clinical isolated strains were drug resistance (73.08％),while PCR-SSCP indicated 65.38％ (32/52) rpoB gene polymorphism. There was no statistic significance(χ2= 2.4914) between the 2 methods. Conclusion:Combined the application of PCR-SSCP with AST in detecting rpoB drug resistant gene polymorphism of M. tuberculosis L-form from pneumoconiosis patients with tuberculosis may have advantages at earlier diagnosis and guidance of clinical medications.展开更多
To study the characteristics of drug-resistant genetic mutation of rpoB on coal workers＇ pneumoconiosis complicated with L-form of Mycobacterium tuberculosis. Methods： A total of 42 clinical isolated strains of Myco...To study the characteristics of drug-resistant genetic mutation of rpoB on coal workers＇ pneumoconiosis complicated with L-form of Mycobacterium tuberculosis. Methods： A total of 42 clinical isolated strains of Mycobacterium tuberculosis L-forms were collected, including 31 drug-resistant strains. Their genomes DNA were extracted, the target genes were amplified by PCR, and the hot regions in the rpoB gene were analyzed by automated DNA sequenator. Results： No mutation of rpoB gene was identified in 11 rifampicin-sensitive strains while conformation changes were found in 31 rifampicin-resistant strains. The mutation rate was 93.55% （29/31） in resistant strains, mainly concentrated in codon 531 （51.6%, 16/31） and 526 （32.26%, 10/31）. Base substitutions happened, including 27 unit point mutation and 2 two point mutation. The mutation of codon 516 that new found wasn＇t reported by internal and overseas scholars. Conclusion： The substitution of highly conserved amino acids encoded by rpoB gene results in the molecular mechanism responsible for rifampicin resistance in Mycobacterium tuberculosis L-forms. It also proves that rpoB gene is diversiform.展开更多
Objective：To study the relationship between mutation in the katG gene and drug resistance of INH in Mycobacterium tuberculosis L-forms among patients of pneumoconiosis complicated with pulmonary tuberculosis, and to ...Objective：To study the relationship between mutation in the katG gene and drug resistance of INH in Mycobacterium tuberculosis L-forms among patients of pneumoconiosis complicated with pulmonary tuberculosis, and to explore the clinical application of PCR-SSCP. Methods： A total of 52 clinical isolated strains of Mycobacterium tuberculosis L-forms were collected. Mutation in the katG genes was detected by PCR-SSCP and traditional antimicrobial susceptibility test （AST）. Results： The results by AST showed that there were 40 persisters in 52 clinical isolated strains. The drug resistant rate was 76.92%（40/52）, and the gene mutation rate of katG was 57.70%（30/52）by PCR-SSCP, the difference was no quite significance （X^2 = 2.8507, P 〉 0.05）. The coincidence rate of two methods was 75.00% （30/40）. Conclusion： The detectionrate of katG resistant strains in Mycobacterium tuberculosis L-forms was high by PCR-SSCP. The combined application of PCR-SSCP and traditional antimicrobial susceptibility test can improve the detecting rate.展开更多
Objective： To study the relationship between drug resistant genetic mutation and drug resistance in Mycobacterium tuberculosis L-form, discuss the internal relationship between drug resistances and drug-resistant rel...Objective： To study the relationship between drug resistant genetic mutation and drug resistance in Mycobacterium tuberculosis L-form, discuss the internal relationship between drug resistances and drug-resistant related genes and explore the value of PCR- SSCP to clinical application. Methods： A total of 52 clinically isolated strains of tuberculosis L-form were collected among 97 pneumoconiosis patients complicated with tuberculosis. The gene mutations of katG, rpoB and rpsL were detected by PCR-SSCP, and the results were compared with those analyzed by traditional antimicrobial susceptibility test（AST）. Results： The gene muta- tion rates of katG, rpoB and rpsL by PCR-SSCP were respectively 57.70% （30/52）, 65.38% （32/52） and 40.38% （21/52）. The rate of reversion was 78.85%（41/52） and the result of drag-resistant genes was invariable. The results of AST showed that there were 40 （76.92%） multi-drug resistant strains in 52 clinically isolated strains. The number for three-drug resistant strain was 21 （40.38%） and that of two-drug resistant was 19（36.54%）, but only 12（23.08%） strains were one drug resistant. The rate of total drug-resistance was 100%, but there were 15 strains of allied mutation of three genes, 16 of two mutations and 6 of only one by PCR-SSCP. The coincidences were respectively 71.43%, 84.12% and 50.00%. Then there was no significant difference between the allied mutations of multi-drug resistant gene and the mutations of only one drug resistant gene （P 〉 0.05）. Conclusion： PCR-SSCP technique has a higher sensibility and specificity to detect the genes of katG, rpoB and rpsL in tuberculosis L-form among pneumoconiosis complicated with tuberculosis,and the detecting rate of two drug resistant strains and three drug resistant strains was higher. The combined application of PCR-SSCP and AST has advantages at earlier diagnosis and guidance of clinical medications.展开更多
在解析数论领域,探讨全纯尖形式的傅里叶系数具有重要的理论价值.通过复变函数的积分技巧,结合自守L-函数的凸界以及积分均值估计方法,研究了不同序列上傅里叶系数的一致均值估计,形式化表达为∑n≤xλf nλf n j,j=2,3,其中f是全模群Γ...在解析数论领域,探讨全纯尖形式的傅里叶系数具有重要的理论价值.通过复变函数的积分技巧,结合自守L-函数的凸界以及积分均值估计方法,研究了不同序列上傅里叶系数的一致均值估计,形式化表达为∑n≤xλf nλf n j,j=2,3,其中f是全模群Γ=SL(2,Z)上权为偶数k的Hecke特征型,λf n是其在尖点∞处傅里叶展开的第n个标准化傅里叶系数.展开更多
文摘In recent years, valuable experience and insights have been gained into L-forms (cell-wall-deficient variants) of bacteria and fungi and their disease-trigger potential in cases with chronic infections, autism spectrum disorders, autoimmune and neurodegenerative diseases. Based on the concept of “internal” blood microbiota, consisting of L-forms and its relevance to health and disease, the current study aims to outline the profile of dysbiotic disorders in three cancer patients (with endometrial cancer, breast cancer and acute myeloid leukemia), all in a phase before chemotherapy. Venous blood samples from the patients and from one control healthy person, were microbiologically studied. The used novel methodology of blood microbiota assessment was based on the following phases: isolation of L-forms, development and propagation, cultivation and conversion of L-forms into classical bacteria and fungi, as well as their identification with MALDI-TOF method. From the patients were isolated L-forms of opportunistic bacteria (Enterococcus faecalis, Esherichia coli, Enterobacter cloacae and Pseudomonas oryzihabitans) and fungi such as Rhodotorula mucilaginosa, Aspergillus fumigatus and Mucorales. In conclusion, the common feature found for the three cancer patients was the isolation from the blood of highly associated communities consisting of morphologically indistinguishable L-bodies, which through reversion in broth, were identified as distinct bacterial and fungal species. Unlike classic bacteria or fungi causing sepsis and bacteremia/fungemia, the presence of L-forms in blood is hidden, it does not demonstrate clinical signs nor it can be detected by conventional methods. It should be noted, however, that the dysbiotic blood microbiota shows unique and individual characteristics for the concrete cancer patient, correlates to the common state of the organism and tumor localization in the body, as well as it outlines the cancer promoting role of L-forms in processes of malignization, cancer genesis and progression.
文摘<b><span style="font-family:Verdana;">Background</span></b><span style="font-family:""><span style="font-family:Verdana;">: The recognition of human blood microbiota, consisting of cell wall-deficient microbes (L-forms), is a major challenge today in the field of microbiology. There are accumulating data confirming the concept of “internal” blood L-form microbiota and its significance for health and diseases. Finding out whether the blood microbiota can be of diagnostic and prognostic importance for detection and evaluation of chronic infections anywhere in </span><span style="font-family:Verdana;">the body is a major objective. In the context of chronically infected upper</span> <span style="font-family:Verdana;">respiratory tract (URT), the aim of the current study was to understand</span><span style="font-family:Verdana;"> wheth</span><span style="font-family:Verdana;">er a local infection can be a source for entry of bacteria and fungi in th</span><span style="font-family:Verdana;">e blood. </span><b><span style="font-family:Verdana;">Methods: </span></b><span style="font-family:Verdana;">Blood samples from six persons with chronic inflammations</span><span style="font-family:Verdana;"> in URT diagnosed with hypertrophied adenoids, chronic sinusitis, nasal polyps, chronic naso-pharyngitis and one control healthy person were studied. Blood microbiota assessment methodology that be used, included three phases: </span></span><span style="font-family:Verdana;">1</span><span style="font-family:""><span style="font-family:Verdana;">) </span><span style="font-family:Verdana;">isolation of L-form cultures from blood-development and propagation;</span></span><span style="font-family:Verdana;">2</span><span style="font-family:""><span style="font-family:Verdana;">) cultivation directed to conversion of L-forms into bacterial and fungal cul</span><span style="font-family:Verdana;">tures;</span></span><span style="font-family:Verdana;">3</span><span style="font-family:Verdana;">) isolation of pure classical bacterial and fungal cultures and their</span><span style="font-family:""> <span><span style="font-family:Verdana;">identification by MALDI-TOF method. </span><b><span style="font-family:Verdana;">Results:</span></b><span style="font-family:Verdana;"> From the patients were isolated </span></span><span style="font-family:Verdana;">L-forms of opportunistic bacteria (</span><i><span style="font-family:Verdana;">Streptococcus mitis</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Roseomonas mucosa</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Dermacoccus nishinomiyaensis</span></i><span><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Enterococcus faecalis</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Acinetobacter johnsonii</span></i><span style="font-family:Verdana;">, </span></span><i><span style="font-family:Verdana;">Pseudomonas putida</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Staphylococcus aureus</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Pseudomonas luteola</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Enterobacter cloacae</span></i><span style="font-family:Verdana;">) and fungi such as </span><i><span style="font-family:Verdana;">Rhodotorula mucilaginosa</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Aspergillus niger</span></i><span style="font-family:Verdana;">,</span><i><span style="font-family:Verdana;"> Aspergillus fumigatus and Mucorales.</span></i> <b><span style="font-family:Verdana;">Conclusion:</span></b><span style="font-family:Verdana;"> The novel innovative methodology for assessment of blood L-form microbiota was successfully applied for detection of microbes responsible for chronic infections at URT.
文摘Objective: To study the relationship between the polymorphism of drug resistant gene rpoB and drug resistance against rifampicin(RFP) of M. tuberculosis L-forms, and to evaluate its clinical application. Methods: A total of 52 clinical isolated strains of M. tuberculosis L-forms were collected. rpoB gene polymorphism was analyzed by polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP) and conventional antimicrobial susceptibility test (AST). Their results were compared. Results: AST results showed that 38 of 52 clinical isolated strains were drug resistance (73.08％),while PCR-SSCP indicated 65.38％ (32/52) rpoB gene polymorphism. There was no statistic significance(χ2= 2.4914) between the 2 methods. Conclusion:Combined the application of PCR-SSCP with AST in detecting rpoB drug resistant gene polymorphism of M. tuberculosis L-form from pneumoconiosis patients with tuberculosis may have advantages at earlier diagnosis and guidance of clinical medications.
基金Supported by the Natural Science Foundation of Universities of Anhui Province (KJ2008A152)the Natural Science Foundation of the Committee of Education of Anhui Province (2005KJ238)
文摘To study the characteristics of drug-resistant genetic mutation of rpoB on coal workers＇ pneumoconiosis complicated with L-form of Mycobacterium tuberculosis. Methods： A total of 42 clinical isolated strains of Mycobacterium tuberculosis L-forms were collected, including 31 drug-resistant strains. Their genomes DNA were extracted, the target genes were amplified by PCR, and the hot regions in the rpoB gene were analyzed by automated DNA sequenator. Results： No mutation of rpoB gene was identified in 11 rifampicin-sensitive strains while conformation changes were found in 31 rifampicin-resistant strains. The mutation rate was 93.55% （29/31） in resistant strains, mainly concentrated in codon 531 （51.6%, 16/31） and 526 （32.26%, 10/31）. Base substitutions happened, including 27 unit point mutation and 2 two point mutation. The mutation of codon 516 that new found wasn＇t reported by internal and overseas scholars. Conclusion： The substitution of highly conserved amino acids encoded by rpoB gene results in the molecular mechanism responsible for rifampicin resistance in Mycobacterium tuberculosis L-forms. It also proves that rpoB gene is diversiform.
基金National Science Foundation Youth fund of Anhui University of Science & Technology(200537)
文摘Objective：To study the relationship between mutation in the katG gene and drug resistance of INH in Mycobacterium tuberculosis L-forms among patients of pneumoconiosis complicated with pulmonary tuberculosis, and to explore the clinical application of PCR-SSCP. Methods： A total of 52 clinical isolated strains of Mycobacterium tuberculosis L-forms were collected. Mutation in the katG genes was detected by PCR-SSCP and traditional antimicrobial susceptibility test （AST）. Results： The results by AST showed that there were 40 persisters in 52 clinical isolated strains. The drug resistant rate was 76.92%（40/52）, and the gene mutation rate of katG was 57.70%（30/52）by PCR-SSCP, the difference was no quite significance （X^2 = 2.8507, P 〉 0.05）. The coincidence rate of two methods was 75.00% （30/40）. Conclusion： The detectionrate of katG resistant strains in Mycobacterium tuberculosis L-forms was high by PCR-SSCP. The combined application of PCR-SSCP and traditional antimicrobial susceptibility test can improve the detecting rate.
基金This work was supported by the Youth Natural Science Foundation of Anhui University of Science & Technology(200537)
文摘Objective： To study the relationship between drug resistant genetic mutation and drug resistance in Mycobacterium tuberculosis L-form, discuss the internal relationship between drug resistances and drug-resistant related genes and explore the value of PCR- SSCP to clinical application. Methods： A total of 52 clinically isolated strains of tuberculosis L-form were collected among 97 pneumoconiosis patients complicated with tuberculosis. The gene mutations of katG, rpoB and rpsL were detected by PCR-SSCP, and the results were compared with those analyzed by traditional antimicrobial susceptibility test（AST）. Results： The gene muta- tion rates of katG, rpoB and rpsL by PCR-SSCP were respectively 57.70% （30/52）, 65.38% （32/52） and 40.38% （21/52）. The rate of reversion was 78.85%（41/52） and the result of drag-resistant genes was invariable. The results of AST showed that there were 40 （76.92%） multi-drug resistant strains in 52 clinically isolated strains. The number for three-drug resistant strain was 21 （40.38%） and that of two-drug resistant was 19（36.54%）, but only 12（23.08%） strains were one drug resistant. The rate of total drug-resistance was 100%, but there were 15 strains of allied mutation of three genes, 16 of two mutations and 6 of only one by PCR-SSCP. The coincidences were respectively 71.43%, 84.12% and 50.00%. Then there was no significant difference between the allied mutations of multi-drug resistant gene and the mutations of only one drug resistant gene （P 〉 0.05）. Conclusion： PCR-SSCP technique has a higher sensibility and specificity to detect the genes of katG, rpoB and rpsL in tuberculosis L-form among pneumoconiosis complicated with tuberculosis,and the detecting rate of two drug resistant strains and three drug resistant strains was higher. The combined application of PCR-SSCP and AST has advantages at earlier diagnosis and guidance of clinical medications.
文摘在解析数论领域,探讨全纯尖形式的傅里叶系数具有重要的理论价值.通过复变函数的积分技巧,结合自守L-函数的凸界以及积分均值估计方法,研究了不同序列上傅里叶系数的一致均值估计,形式化表达为∑n≤xλf nλf n j,j=2,3,其中f是全模群Γ=SL(2,Z)上权为偶数k的Hecke特征型,λf n是其在尖点∞处傅里叶展开的第n个标准化傅里叶系数.
