Empirical Study On Relations Between Nanobacteria And Type Ⅲ Prostatitis As Well As Prostatic Calculus

Background and Objectives TypeⅢprostatitis (CPPS) is the most common Chronic prostatitis,with uncertain etiological factor and extremely difficult treatment. Surveys and numerous hypothesis on the cause of the infection haven been proposed, yet the clinical characteristics of the disease could not be thoroughly elucidated. The existence of calcification foci and the significant therapeutic effect of antibiotic therapy indicate the possible existence of infective factor that could not be detected through routine examination methods.Calculus could be usually found in the prostate of patients with CPPS,and such prostatic calculus was a kind of true calculus formed in gland alveolus and glandular tube of prostate with uncertain factors for its formation including chronic inflammation of prostate gland alveolus and excretory duct, functions of repressive or promoting nucleating factor, concentration of calcium ion. It is worth exploring whether some unknown infective etiological factor was conductive to the formation of prostatic calculus.The etiological factor of CPPS as well as prostatic calculus could be surveyed by a brand new means with the discovery of nanobacteria and its etiological characteristic of the bacteria.A newly-discovered nano-scale (50～200nm) coccobacteria, nanobacteria could not grow under the common microbial culture condition. Infection of various kinds of human cells and extraskeletal calcification (pathologic calcification) of the infected cells could be caused by the bacteria. Simutaneously toxins leading to tissue inflammation were released. The study on mechanism of TypeⅢprostatitis and prostatic calculus was thus provided with a new perspective with the discovery. It pays to further explore the field since pertinent literature and report have not been available yet. The etiological effect of nanobacteria on CPPS and prostatic calculus was expected to be approved through the study, in addition, the therapia treating CPPS and prostatic calculus by means of eliminating nanobacteria was expected to gain theoretical evidence in the study.Methods Nanobacteria were arranged to be fractionally cultivated from the large sample of secretory juice of typeⅢprostatitis and prostatic calculus. Methods including indirect immunofluorescence, calcifying-nodule staining, electron microscope observation, and 16s rRNA gene amplification and sequencing were applied to the identification of the suspicious culture. The etiological effect of nanobacteria on CPPS and prostatic calculus was studied to provide a new etiological basis for the clinical treatment.Results1,After 3-4 weeks , we can find white precipitate adhering to the tube in part of specimens.2,As high as 58 % positive rate of nanobacteria was obtained in the group of patients with CPPS, while the positive rate in the normal control was only 4 %. The difference of the positive rate had statistical significance(χ2 =34.08 P <0.01).3,The indirect IF staining of monoclonal antibody was applied to the identification of nanobacteria, which were found in 26 samples of prostatic calculus with the positive rate of 65%. The positive rate of nanobacteria was 83.3 % with white precipitate adhering to the tube ,and was 37.5 % with flocculent precipitate or without precipitate (χ2 =8.86 P < 0.01) .4,By negative staining ,the sizes of nanobacteria ranged from 100 to 500 nm, appeared coccoid- coccobacillary in shape .5,By calcific and Gram staining, nanobacteria distributed with clustering.6,The resemblance of specificity fragment and 16SrRNA Gene of nanobacteria was 98%:Score = 2480 bits (1290), Expect = 0.0, Identities = 1387/1409 (98%), Gaps = 4/1409 (0%), Strand=Plus/Plus.Conclusion and Prospect1,The indirect IF staining of monoclonal antibody was applied to the identification of nanobacteria. As high as 58 % positive rate of nanobacteria was obtained in the group of patients with CPPS, while the positive rate in the normal control was only 4 %. The difference of the positive rate had statistical significance(χ2 =34.08 P <0.01), which approved the close correlation between morbility of typeⅢprostatitis and nanobacterial infection. 2,The indirect IF staining of monoclonal antibody was applied to the identification of nanobacteria, which were found in 26 samples of prostatic calculus with the positive rate of 65%. The close correlation between morbility of prostatic calculus and nanobacterial infection was also approved.3,The Gene of example were identified by the PCR method . PCR products were analyzed by agarose gel electrophoresis and sequencing . PCR and Sequencing are useful inidentifying nanobacteria.4,Morphology identification was time-consuming considering the long culture period of nanobacteria, while PCR method, vector T cloning and sequencing were convenient methods for identification of 16SrRNA gene of nanobacteria. An expeditious detecting method for the identification of nanobacteria was thus found, and the research basis for the mineralized gene of nanobacteria was established.5,Nanobacteria might be a microbe playing an important role in the morbility of CPPS and prostatic calculus. A parallel might be drawn between their correlation and relation between HP and gastric ulcer. Clinically speaking, CPPS and prostatic calculus could be treated through eliminating nanobacteria in accordance with the microbial characteristics of nanobacteria. Thus a trail was blazed in the field of radical cure of CPPS and prostatic calculus.