【作者】 彭风华；

【导师】 杨金瑞；

【作者基本信息】 中南大学 ， 外科学， 2006， 博士

【摘要】 第一部分前列腺炎样症状患者转归调查目的调查“前列腺炎样症状”患者的转归预后及影响临床疗效相关因素，评估当前“慢性前列腺炎”患者的诊疗效果，随访并根据疗效筛选出典型的顽固性慢性前列腺炎患者参与第二部分实验室研究。方法2004年6月～2004年9月泌尿外科门诊144例“前列腺炎样症状”患者入选，均采用美国国立卫生院慢性前列腺炎症状问卷(NIH-CPSI)调查，取按摩后前列腺液行前列腺液常规检查。患者以按摩后前列腺液白细胞数目为依据，按NIH1995标准分类，接受推荐方案治疗，分别于第6周及1年后接受随访。结果在规律治疗6周后，总计115例患者继续参与研究。115例前列腺炎样症状患者痊愈率只有13.9％，51.3％患者获得了不同程度的症状缓解或好转，34.8％的患者症状无改善或病情加重。前列腺炎样症状转归与按摩后前列腺液白细胞数目、年龄、有无支原体或衣原体、病程长短等无关。随访1年62例患者继续参与研究，痊愈率只有19.3％，46.7％患者获得了不同程度的症状改善，而33.9％的患者症状无改善或病情加重。ⅢA与ⅢB型前列腺炎样症状患者在治疗过程中疾病类型可以相互转换，由ⅢA型向ⅢB型转换较为多见，由ⅢB型向ⅢA型转换较为少见。结论采用NIH-CPSI问卷调查结合临床基本检查-前列腺液常规检查、细菌培养，有助于临床前列腺炎的诊断，但前列腺炎样症状其治疗在临床上仍然是一大难题，好转痊愈率较低。以前列腺液白细胞计数为分类基础的前列腺炎样症状各种类型之间在自然病程过程中，存在相互转换，这一结果在对慢性前列腺炎科研数据进行分析时有着重要意义。第二部分细胞因子及其受体基因多态性与慢性前列腺炎／慢性盆腔疼痛综合症的关系目的了解细胞因子及其受体基因多态性在慢性前列腺炎／慢性盆腔疼痛综合症组和正常对照组中的分布，探讨其在慢性前列腺炎发病中的作用及其在诊断及治疗中的意义。方法正常对照组51人，从第一部分研究中筛选出典型CP／CPPS患者24人，均为湖南汉族。采用TBG公司DNA10分钟快速提取试剂盒提取DNA，使用美国PEL-FREEZ公司的细胞因子基因型检测试剂盒，采用序列特异性引物聚合酶链反应方法(SSP-PCR)检测受试者13种细胞因子和细胞因子受体22个位点的基因型：IL-1α-889C／T、IL-1β(-511C／T、+3962C／T)、IL-1RPstⅠ970 C／T、IL-1Ra mspⅠ11100C／T、IL-4Rα+1902A／G、IL-12-1188A／C、IFN-γUTR5644A／T、TGF-β1(密码子10C／T、密码子25C／G)、TNF-α(-308A／G、-238A／G)、IL-2(-330T／G、+166G／T)、IL-4(-1098G／T、-590C／T、-33C／T)、IL-6(-174C／G、nt565A／G)、IL-10(-1082A／G、-819C／T、-592A／C)；采用双抗体夹心ELISA法，检测51例正常对照者外周血IL-10含量，同时检测20例CP／CPPS患者按摩后前列腺液IL-10含量。结果1．汉族人群在IL-10-1082位点，G等位基因频率(在患者人群2.1％，在正常对照组4.9％)，明显低于A等位基因频率(在患者人群97.9％，在正常对照组95.1％)且该位点基因型在两组间分布无显著性差别。2．和对照组比较，患者组在IL-10(-819C／T)位点T／T基因型(62.5％vs 31.3％)更为多见，差别有显著性意义(P＜0.05)；在IL-10(-592C／A)位点A／A基因型(62.5％vs 31.3％)更为多见，差别有显著性意义(P＜0.05)。3．与对照组比较，患者组在-819位点T等位基因频率(75.0％vs55.9％)更为多见，差别有显著性意义(P＜0.05)；在-592位点A等位基因(75.0％vs 55.9％)更为多见，差别有显著性意义(P＜0.05)。4．IL-10(-1082，-819，-592)ATA／ATA基因型在CP／CPPS组较之正常组更为多见(62.5％vs 31.2％，P＜0.05)，差别有统计学意义，与CP／CPPS发病有关。5．IL-10(-1082，-819，-592)的基因型分布，与CP／CPPS患者的年龄、病程、CP／CPPS类型(ⅢA／ⅢB)、CPSI-NIH症状问卷中疼痛项目评分、疼痛程度、射精后痛、排尿症状、排尿痛、客观症状、生活质量影响等各项评分以及总分等均没有显著关系。6．正常对照人群IL-10 ACC／ACC组、GCC／ATA+GCC／ACC组与ATA／ATA组比较，外周血IL-10产量均有明显差别(分别为256.8±209.7 pg／ml，355.2±298.4 pg／ml，110.7±102.6 pg／ml)差别有显著性意义；ATA／ACC组与ATA／ATA组比较，IL-10产量虽有增加(分别为205.1±196.0 pg／m，110.7±102.6 pg／m)但差别无显著性意义(P)0.05)。7．CP／CPPS患者IL-10基因型多态性与前列腺液IL-10浓度无明显关系。8．IL-4-590 T／C位点两组间基因型频率存在组间差别，与对照组比较，患者组T／C杂合子基因型更为多见(50.0％vs 27.5％)，但统计分析其P值临近显著性水平(P=0.056)，差别无统计学显著性意义。9．其他位点基因型频率在两组间没有显著性差异。10．TGFβ(Codon10，25)及TNFα(-308，-238)单倍体型、基因型在两组间频率分布无显著性差异，与CP／CPPS发病关系不大。结论汉族人群抗炎细胞因子基因型多态性与CP／CPPS发病有关，促炎细胞因子及其受体基因多态性与CP／CPPS发病关系不大。IL-10基因多态性决定外周血IL-10产量，CP／CPPS患者中IL-10低产量基因型频率多见。汉族人群中IL-10-1082位点G等位基因频率明显低于白种高加索人。汉族人群IL-10基因与CP／CPPS发病的关系不是表现在IL-10-1082位点基因多态性，而是表现在与IL-10-1082位点相连锁的-819，-592位点基因多态性。IL-10基因多态性并不能直接决定患者前列腺液IL-10含量。分析患者细胞因子基因型多态性有助于采取以人为本的个性化治疗。更多还原

【Abstract】 PART 1 Follow up of patients with "prostatitis-like symptom"Objective To study the clinical curative effect and prognosis of "prostatitis-like symptom" and the relevant factors affecting the curative results. To select the typical stubbornness chronic prostatitis patient to participate in the second part of laboratory research according to curative effect.Methods 144 cases of "prostatitis-like symptom" patients in our hospital were adopted from June, 2004 to September, 2004. All the patients received the investigation based on NIH-CPSI questionnaire. Expressed prostatic fluids(EPS) were acquired and prostatic fluid routine inspection were executed. Patients were classified based on white blood cell count in EPS according to the NIH1995 standard classification, and accepts the recommendated treatment. Patients received follow-up visit after 6 weeks and 1 year respectively.Results After 6 week regular treatment, 115 patients continued to participate in the research. Convalescing rate of the 115 "prostatitis-like symptom" patients was only 13.9%. 51.3% patients obtained the varying degree of alleviation of symptoms or the condition changed for the better.34.8% patients’ symptoms did not have a improvement or the condition aggravated. Curative effect of "prostatitis-like symptom" had nothing to do with the age, the course of illness, neither with whether there was the mycoplasma or the chlamydia as well as white blood cell number in EPS.ⅢA Type andⅢB Type "prostatitis-like symptom" may had the interconversion during patient’s course of treatment. Situation was common byⅢA change to theⅢB, and less wasⅢB to theⅢA.Conclusions Combining the NIH-CPSI questionnaire survey with clinical basic inspections-prostate gland fluid routine inspection and the bacilli culture help to make a clinical diagnosis of prostatitis. "prostatitis-like symptom" was still a difficult problem in clinical aspect, with the amelioration rate and convalescing rate was low. Interconversion existed between two types of "prostatitis-like symptom" during the course of disease, which was meaningful in the case of analyzing scientific research data about chronic prostatitis. PART 2 Association of gene polymorphisms of cytokine and cytokine receptor with chronic prostatitis/chronic pelvic pain syndromeObjective: To research the distribution of gene polymorphisms of cytokines and cytokine receptors in CP/CPPS patients group and health control group. To explore the role of gene polymorphism of cytokine and cytokine receptor in CP/CPPS and their role in diagnosis and treatment of CP/CPPS.Methods: We have genotyped 24 patients diagnosed with typical CP/CPPS which selected from research of part one and 51 health controls from the Han nationality population in Hunan province for the single nucleotide polymorphisms of 13 cytokines at 22 site: IL-lα-889 C/T、IL-1β(-511 C/T、＋3962C/T)、IL-1RPst 1970 C/T, IL-1Ra mspⅠ11100 C/T、IL-4Rα＋1902 A/G、IL-12-1188 A/C、IFN-γUTR5644 A/T、TGF-β1 (codon10 C/T、codon25 C/G)、TNF-α(-308 A/G、-238 A/G)、IL-2 (-330 T/G、＋166G/T)、IL-4 (-1098 G/T、-590 C/T、-33C/T)、IL-6 (-174 C/ G、nt565 A/G)、IL-10 (-1082 A/ G、-819C/T、-592 A/C). Clinical data were collected on all patients. DNA was extracted with BioGene-Expuze 10 DNA kit supplied by Texas BioGene Incorporation. Genotyping was carried out using SSP-PCR technique with cytokine genetyping kit supplied by PEL-FREEZ Incorporation. IL-10 level in peripheral blood of 51 health controls and in EPS of 20 CP/CPPS patients were tested using ELISA technique.Results:1.The frequency of G allele (2.1% in patients group, 4.9% in health control group) of IL-10 at -1082 was lower than that of A allele(97.9% in patients group, 95.1% in health control group) obviously in Han nationality population. There were no significant difference of allele frequencies between patients group and health control group.2.A statistically significant difference was observed in the genetype frequency of the polymorphisms between the group of patients and the group of healthy individuals with the polymorphism of the IL-10 at -1082,where frequency of T/T genetype was higher in patient group(62.5% vs 31.3%,p＜0.05). A statistically significant difference was observed in the genetype frequency of the polymorphisms between the group of patients and the group of healthy individuals with the polymorphism of the IL-10 at -592, where frequency of A/A genetype was higher in patient group(62.5% vs 31.3%,p＜0.05).3.A statistically significant difference was observed in the allele frequency of the polymorphisms between the group of patients and the group of healthy individuals with the polymorphism of the IL-10 at -819, where frequency of T allele was higher in patient group(75.0% vs 55.9%,p＜0.05). A statistically significant difference was observed in the allele frequency of the polymorphisms between the group of patients and the group of healthy individuals with the polymorphism of the IL-10 at -592, where frequency of A allele was higher in patient group(75.0% vs 55.9%,p＜0.05).4.A statistically significant difference was observed in the genetype frequency of the polymorphisms between the group of patients and the group of healthy individuals with the polymorphism of the IL-10 at -1082,-819,-592, where frequency of ATA/ATA genetype was higher in patient group (62.5% vs 31.2%,P＜0.05 )5.There were no statistically significant association of distribution of genetype frequency with age, course, CP/CPPStype (ⅢA/ⅢB) and neither with score of pain, pain degree, pain after ejaculation, symptom with void, pain during void, objective symptom, quality of life according to NIH-CPSI questionnaire.6.The IL-10 level in peripheral blood of ACC/ACC group and GCC/ATA＋GCC/ACC group were statistically significantly different from that of ATA/ATA group(256.8±209.7 pg/ml, 355.2±298.4 pg/ml, 110.7±102.6 pg/ml,respectively). Level of IL-10 in ATA/ACC group was higher than that in ATA/ATA group, however the difference was not significant.7.Gene polymorphism at -1082,-819,-592 of IL-10 have no statistically significant association with levels of IL-10 in EPS of CP/CPPS patients. 8. Genetype T/C of IL-4 at -590 was found more frequently in the patients (50.0% vs 27.5%), while the difference was not statistically significant,with P value was 0.056 that was cloes to the critical value.9. There was no statistically significant difference observed between healthy group and patients group on other researched gene polymorphisms.10. No statistically significant difference was observed in the haplotype frequency and genetype frequency of TGFβgene (codon 10, condon 25) and TNFαgene (-308, -238) between the CP/CPPS patients and the healthy control group.Conclusions: Our data suggested that anti-inflammation cytokine gene polymorphisms but not proinflammation cytokine gene polymorphisms were associated with the pathogenesis of CP/CPPS. IL-10 gene polymorphism prearranged the level of IL-10 in peripheral blood. Low product genetype of IL-10 was more frequently obserbed in CP/CPPS population. Frequency of allele G of IL-10 at -1082 was significantly lower in Han nationality population than that in Caucasians. Gene polymorphisms of IL-10 at -1082 cann’t establish its role in pathogenesis of CP/CPPS, however gene polymorphisms of IL- 10 at -819 and at -592, which were linkage with site -1082, can play its role in etiology of CP/CPPS. Gene polymorphism of IL-10 cann’t prearrange the level of IL-10 in EPS of CP/CPPS patients. Analysis of gene polyphisms of cytokine help the patient to accepte a individual treatment in consideration of humanity.更多还原