Purpose: Identifying the source of stone formation in recurrent stone formers has always been a big problem. Material and Methods: In this study kidney stones from 52 patients were submitted to direct examination by s...Purpose: Identifying the source of stone formation in recurrent stone formers has always been a big problem. Material and Methods: In this study kidney stones from 52 patients were submitted to direct examination by scanning electronic microscopy (SEM) after manual fracture and 27 calculi were cultured in Eagle’s Minimum Essential Medium (E-MEM) and Brain-Heart Infusion (BHI) for eight weeks at 37°C in 5% CO2 atmosphere. Twenty-seven powdered and demineralized stones were suspended in sterile PBS, filtered through 0.22 m-pore-size sterile filters Millex (Millipore, Massachusetts, USA) and submitted to DNA extraction (Quiagen-Brazil). Platinum PCR SuperMIX (GIBCO-BRL), primers (Invitrogen), and Ultra Pure Water (Advanced Biotechnologies, Columbia, MD) were used for PCR (Polymerase Chain Reaction), which was generally conducted for 30 or 35 cycles with annealing of primers for 40 sec at 55°C and extension for 1 min at 72°C. Results: In 36 out 52 (69%) kidney stones it was detected the presence of biofilm coating the mineral surface of the stone when examined by SEM, consisting of coccoid particles, isolated or clustered, with diameter of 500 nm or less. Eleven out 27 (41%) kidney stone cultures produced white-colored sediment on the bottom of the tubes after eight-week incubation, revealing tiny structures similar to those observed directly by SEM. These structures were similar in size and morphology to spherical apatite particles previously observed in human kidney stones and named as nanobacteria (NB). No PCR products were observed in the samples. Conclusion: We found a strong correlation between renal stones and calcifying nanoparticles (CNP) in this study and these results open a new insight on this area to explore the etiology of stone formation. Whether NB/CNP are truly microorganisms or self-propagating mineral compounds is still controversial and its contribution, if any, in apatite nucleation and crystal growth remains uncertain.展开更多
Objective: Analysis of the effect of nanobacteria on the expression of local biomarkers in patients with rheumatoid arthritis. Methods: Serum samples from 54 patients with rheumatoid arthritis in our hospital were tes...Objective: Analysis of the effect of nanobacteria on the expression of local biomarkers in patients with rheumatoid arthritis. Methods: Serum samples from 54 patients with rheumatoid arthritis in our hospital were tested for nanobacteria, and were divided into three groups based on the concentration of nanobacteria. To compare the differences of main essential elements, serum cytokines, immunochemical indicators and bone metabolic markers in synovial fluid of three groups of patients. Results: Differences in serum calcium levels between the three groups are significant. The differences in IL-1, TNF-α and CRP were significant. The difference of IgM, IgA, anti-CCPP content are significant. CTX-1, RF, ESR content differences are significant. Conclusions: The detection of nanobacteria in joint fluid of patients with rheumatoid arthritis has certain effects on the expression of serum cytokines and immune indexes. The higher the concentration of nanobacteria, the higher the expression of inflammatory and immunological indicators in serum.展开更多
In 1990, nanobacterium was found and named by Kajander.~1 With distinct mineralizing ability, nanobacteria are thought to play a role in extraskeletal calcifying diseases. It have been found in many human tissues, but...In 1990, nanobacterium was found and named by Kajander.~1 With distinct mineralizing ability, nanobacteria are thought to play a role in extraskeletal calcifying diseases. It have been found in many human tissues, but whether they exist in the bile or gallbladder mucosa remains unclear. The present study was undertaken to investigate by ELISA, bacterial culturing, immunohistochemical staining and transmission electron microscopy (TEM), whether nanobacteria exist in serum, bile or gallbladder mucosa of healthy people and patients with cholecystolithiasis.展开更多
目的探讨乌梅提取物对纳米细菌(NB)导致的大鼠肾草酸钙结石形成的影响及其机制。方法 30只大鼠随机分为A、B、C、D、E组各6只。A组第1天以生理盐水1 m L尾静脉一次性注射作为正常对照,B、C、D、E组均于第1天以尾静脉一次性注射NB混悬液1...目的探讨乌梅提取物对纳米细菌(NB)导致的大鼠肾草酸钙结石形成的影响及其机制。方法 30只大鼠随机分为A、B、C、D、E组各6只。A组第1天以生理盐水1 m L尾静脉一次性注射作为正常对照,B、C、D、E组均于第1天以尾静脉一次性注射NB混悬液1 m L建立大鼠草酸钙结石模型。A、B组第2天起均以生理盐水2m L/d灌胃,C、D、E组第2天起分别以0.05、0.10、0.15 g/m L乌梅提取物2 m L/d灌胃。各组干预4周后,收集24 h尿液,应用高锰酸钾褪色法测定尿液草酸(Ox)水平,全自动生化仪测定尿液Ca^(2+)水平,柠檬酸试剂盒测定尿液柠檬酸水平,8-羟基脱氧鸟苷(8-OHDG)测试盒测定尿液8-OHDG水平。取左侧肾脏,行HE染色观察肾组织草酸钙结晶形态,行结晶量分级评分。采用硫代巴比妥酸法检测肾组织丙二醛(MDA)水平。结果 B组尿液Ox、Ca^(2+)、8-OHDG水平均高于A组,柠檬酸水平低于A组(P均<0.05);C、D、E组尿液Ox、Ca^(2+)、8-OHDG水平均低于B组,柠檬酸水平均高于B组(P均<0.05)。A组无草酸钙结晶,B组草酸钙结晶明显较多,C、D、E组草酸钙结晶数量较B组明显减少。B组草酸钙结晶量评分高于A组,C、D、E组草酸钙结晶量评分均低于B组(P均<0.05)。B组肾组织MDA水平高于A组,D、E组肾组织MDA水平均低于B组(P均<0.05)。结论乌梅提取物对NB所致的大鼠肾草酸钙结石具有明显的抑制作用,其机制可能与降低大鼠尿液Ox水平,进而减少尿Ca^(2+)排泄、减轻脂质过氧化损伤有关。展开更多
目的构建纳米细菌(NB)致肾结石大鼠模型,观察乌梅提取物对早期肾损伤分子1(KIM-1)和骨桥蛋白(OPN)的影响,分析乌梅提取物对早期肾结石的治疗意义。方法从人上尿路结石分离并培养NB。将54只雄性SD大鼠随机分为空白组、诱石组和治疗组,采...目的构建纳米细菌(NB)致肾结石大鼠模型,观察乌梅提取物对早期肾损伤分子1(KIM-1)和骨桥蛋白(OPN)的影响,分析乌梅提取物对早期肾结石的治疗意义。方法从人上尿路结石分离并培养NB。将54只雄性SD大鼠随机分为空白组、诱石组和治疗组,采取尾静脉注射NB构建肾结石模型,以乌梅提取物干预模型,并在不同时期分批处死大鼠,实时PCR检测肾组织KIM-1 m RNA的表达,ELISA检测尿KIM-1浓度,HE染色观察肾组织结石晶体情况,并用免疫组化方法检测OPN表达。结果诱石组及治疗组肾结石模型大鼠出现肾小管扩张,肾小管结石晶体形成,早期KIM-1和OPN的表达升高,上述变化与NB的注射时间呈正相关,乌梅提取物可拮抗该类变化。结论乌梅提取物可能通过修复肾损伤减少肾结石形成,该机制可能与乌梅提取物对KIM-1、OPN的基因调节有关。展开更多
文摘Purpose: Identifying the source of stone formation in recurrent stone formers has always been a big problem. Material and Methods: In this study kidney stones from 52 patients were submitted to direct examination by scanning electronic microscopy (SEM) after manual fracture and 27 calculi were cultured in Eagle’s Minimum Essential Medium (E-MEM) and Brain-Heart Infusion (BHI) for eight weeks at 37°C in 5% CO2 atmosphere. Twenty-seven powdered and demineralized stones were suspended in sterile PBS, filtered through 0.22 m-pore-size sterile filters Millex (Millipore, Massachusetts, USA) and submitted to DNA extraction (Quiagen-Brazil). Platinum PCR SuperMIX (GIBCO-BRL), primers (Invitrogen), and Ultra Pure Water (Advanced Biotechnologies, Columbia, MD) were used for PCR (Polymerase Chain Reaction), which was generally conducted for 30 or 35 cycles with annealing of primers for 40 sec at 55°C and extension for 1 min at 72°C. Results: In 36 out 52 (69%) kidney stones it was detected the presence of biofilm coating the mineral surface of the stone when examined by SEM, consisting of coccoid particles, isolated or clustered, with diameter of 500 nm or less. Eleven out 27 (41%) kidney stone cultures produced white-colored sediment on the bottom of the tubes after eight-week incubation, revealing tiny structures similar to those observed directly by SEM. These structures were similar in size and morphology to spherical apatite particles previously observed in human kidney stones and named as nanobacteria (NB). No PCR products were observed in the samples. Conclusion: We found a strong correlation between renal stones and calcifying nanoparticles (CNP) in this study and these results open a new insight on this area to explore the etiology of stone formation. Whether NB/CNP are truly microorganisms or self-propagating mineral compounds is still controversial and its contribution, if any, in apatite nucleation and crystal growth remains uncertain.
文摘Objective: Analysis of the effect of nanobacteria on the expression of local biomarkers in patients with rheumatoid arthritis. Methods: Serum samples from 54 patients with rheumatoid arthritis in our hospital were tested for nanobacteria, and were divided into three groups based on the concentration of nanobacteria. To compare the differences of main essential elements, serum cytokines, immunochemical indicators and bone metabolic markers in synovial fluid of three groups of patients. Results: Differences in serum calcium levels between the three groups are significant. The differences in IL-1, TNF-α and CRP were significant. The difference of IgM, IgA, anti-CCPP content are significant. CTX-1, RF, ESR content differences are significant. Conclusions: The detection of nanobacteria in joint fluid of patients with rheumatoid arthritis has certain effects on the expression of serum cytokines and immune indexes. The higher the concentration of nanobacteria, the higher the expression of inflammatory and immunological indicators in serum.
文摘In 1990, nanobacterium was found and named by Kajander.~1 With distinct mineralizing ability, nanobacteria are thought to play a role in extraskeletal calcifying diseases. It have been found in many human tissues, but whether they exist in the bile or gallbladder mucosa remains unclear. The present study was undertaken to investigate by ELISA, bacterial culturing, immunohistochemical staining and transmission electron microscopy (TEM), whether nanobacteria exist in serum, bile or gallbladder mucosa of healthy people and patients with cholecystolithiasis.
文摘目的探讨乌梅提取物对纳米细菌(NB)导致的大鼠肾草酸钙结石形成的影响及其机制。方法 30只大鼠随机分为A、B、C、D、E组各6只。A组第1天以生理盐水1 m L尾静脉一次性注射作为正常对照,B、C、D、E组均于第1天以尾静脉一次性注射NB混悬液1 m L建立大鼠草酸钙结石模型。A、B组第2天起均以生理盐水2m L/d灌胃,C、D、E组第2天起分别以0.05、0.10、0.15 g/m L乌梅提取物2 m L/d灌胃。各组干预4周后,收集24 h尿液,应用高锰酸钾褪色法测定尿液草酸(Ox)水平,全自动生化仪测定尿液Ca^(2+)水平,柠檬酸试剂盒测定尿液柠檬酸水平,8-羟基脱氧鸟苷(8-OHDG)测试盒测定尿液8-OHDG水平。取左侧肾脏,行HE染色观察肾组织草酸钙结晶形态,行结晶量分级评分。采用硫代巴比妥酸法检测肾组织丙二醛(MDA)水平。结果 B组尿液Ox、Ca^(2+)、8-OHDG水平均高于A组,柠檬酸水平低于A组(P均<0.05);C、D、E组尿液Ox、Ca^(2+)、8-OHDG水平均低于B组,柠檬酸水平均高于B组(P均<0.05)。A组无草酸钙结晶,B组草酸钙结晶明显较多,C、D、E组草酸钙结晶数量较B组明显减少。B组草酸钙结晶量评分高于A组,C、D、E组草酸钙结晶量评分均低于B组(P均<0.05)。B组肾组织MDA水平高于A组,D、E组肾组织MDA水平均低于B组(P均<0.05)。结论乌梅提取物对NB所致的大鼠肾草酸钙结石具有明显的抑制作用,其机制可能与降低大鼠尿液Ox水平,进而减少尿Ca^(2+)排泄、减轻脂质过氧化损伤有关。
文摘目的构建纳米细菌(NB)致肾结石大鼠模型,观察乌梅提取物对早期肾损伤分子1(KIM-1)和骨桥蛋白(OPN)的影响,分析乌梅提取物对早期肾结石的治疗意义。方法从人上尿路结石分离并培养NB。将54只雄性SD大鼠随机分为空白组、诱石组和治疗组,采取尾静脉注射NB构建肾结石模型,以乌梅提取物干预模型,并在不同时期分批处死大鼠,实时PCR检测肾组织KIM-1 m RNA的表达,ELISA检测尿KIM-1浓度,HE染色观察肾组织结石晶体情况,并用免疫组化方法检测OPN表达。结果诱石组及治疗组肾结石模型大鼠出现肾小管扩张,肾小管结石晶体形成,早期KIM-1和OPN的表达升高,上述变化与NB的注射时间呈正相关,乌梅提取物可拮抗该类变化。结论乌梅提取物可能通过修复肾损伤减少肾结石形成,该机制可能与乌梅提取物对KIM-1、OPN的基因调节有关。
