| Background and objectives There have been four types of prostatitis among which type III prostatitis stands as the most challenging one due to its hitherto miserable causes. Although efforts have been made in the past decades to make clear what remains the final draw, no bacteria were found to be the etiological factor. Some investigators have turned to other pathogeny, beyond the bacteria. Unfortunately, all those research results could not explain the clinical characteristics of type III prostatitis well. Therefore, it is urgent to make sure what is the etiological factor that leads to type III prostatitis.Some interesting phenomena were noted in clinical studies. Lots of inflammatory markers are positive in the expressed prostatic secretion(EPS) of most type III prostatitis patients and anti-inflammation is helpful to type III prostatitis. Therefore, we hypothesize that there is a bacterium in prostates of type III prostatitis patients. Moreover, some other evidences also indicate there are unknown bacteria in type III prostatitis patients. For example, there are calcification foci and inflammation in prostate, which might be related with the infection. Such prostatic calculus is a kind of true calculus formed in gland alveolus and glandular tube of prostateNanobacteria(NB) is a new-discovered coccobacteria, which can not grow in the common microbial culture medium, but can grow in cell culture medium. NB can not be detected by routine detective methods, which cause inflammation by releasing toxin and lead extraskeletal calcification (pathologic calcification) in the infected cells. Animal experiments show that NB cause calcification in kidney and gall bladder. NB had been isolated and cultured from EPS of patients with type III prostatitis and prostatic calculus. More importantly, anti-NB therapy can significantly relieve the clinical symptoms of those patients, the stone decrease in size or disappear finally. Based on above-mentioned results, we hypothesize that NB might be a bacterium that causes type III prostatitis and prostatic calculus. In this study, in the first step, we tried to isolate NB from EPS of patients with type III prostatitis, and then observed the pathological changes of prostate in animal after infection of NB. The differential effect of NB, bacillus coli, and pelvic gore on prostates will be compared in this study.Methods1. NB were isolated from EPS of patients with type III prostatitis and prostatic calculus, identified by immunofluorescence, calcifying-nodule staining, electron microscope, 16SrRNA gene amplification and sequencing.2. Adult male SD rats were randomly divided into control group, model group and treatment group respectively. The rats in the model group and treatment group were infused with NB suspension via urethra, and the control rats were infused with normal saline. Those rats in the treatment group were administrated with tetracycline for 4 more weeks after infused with NB 4 weeks. After sacrificed, some parameters were checked, including the prostatic pathological change, body weight, tissue WBC number, lecithine cytorrhyctes, the level of cytokines (IL-1β, TNF-αand IL-6), and NB was re-isolated from the prostates.3. After establishment of models of NB, bacillus coli infection, and pelvic gore, the above-mentioned parameters were checked at 4 and 8 weeks.Results1. Twenty-six of 55 patients with positive NB infection were diagnosed by immunoflurescence, and only 2 in the control group. Respectively, the positive rate was 48.3%, 5%. The difference of the positive rate had statistical significance. Among the 30 samples with prostate calculus, NB can be detected in17 samples (56.7%).2. By negative staining, the sizes of NB ranged from 100 to500nm, appeared coccoid-coccobacillary in shape and around with black-like substance.3. Calcific and Gram staining showed NB distributed with clustering.4. The resemblance of specificity fragment and 16SrRNA Gene of NB was 98%.5. The chronic inflammation changes of prostates were observed in model group after infection with NB for 4 weeks; after observation for 8 weeks, the infection still existed, and the prostates tissue was with degeneration and calculus. However, in the treatment group, no calculus was found.6. Although no difference for body weight among all rats, the parameters in infection rats, including prostatic pathological change, body weight, tissue WBC number, lecithine cytorrhyctes and the level of cytokines (IL-1β, TNF-αand IL-6), were significantly abnormal as compared with control rats. After treatment with medicine, all the aberrant parameters were normalized.7. NB was found in all the rat prostate samples of model group after culture, but not in control group.8. Compared with control group, E.coli group and NB group showed a serious chronic inflammation in prostate tissue after infection for 4 weeks, and the degree of chronic inflammation in E.coli group was more serious than NB group. After 8 weeks, all three groups were observed with chronic inflammation changes except of control group. Moreover, calculus was found only in NB group.9. There was no significant difference of rat weights among four groups. The prostate weights in the E.coli group and NB group were lower. In contrast, the number of WBC and lecithine cytorrhyctes and the level of cytokines (IL-1β, TNF-αand IL-6) were higher than that in control group and pelvic gore group (P<0.05). At the eight week points, there were no significant difference among E.coli group, NB group and pelvic gore group; as compared with control group, the prostate weights of control group were lower, whereas, the count of WBC and lecithine cytorrhyctes and the level of cytokines (IL-1β, TNF-αand IL-6) of the other three groups were higher.Conclusion and Prospect1. NB could be detected in patients with type III prostatitis and prostatic calculus. The positive rate of NB in the patients and prostatic calculus is higher, indicating the relationship between NB infection and type III prostatitis and prostatic calculus.2. NB leads to chronic prostatitis; tetracycline is an effective method to reduce the inflammation of prostatitis.3. NB leads to prostatic calculus formation, which could be blocked in the presence of tetracycline.4. NB could cause the prostatitis, and the prostatitis, induced by NB, is more serious than pelvic gore, but lower than E.coli. Only NB can lead to prostatic calculus formation.5. NB might play an important role in type III prostatitis and prostatic calculus formation, which might provide an alternative method for treatment of type III prostatitis and prostatic calculus. |
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