| Background:Benign prostatic hyperplasia (BPH) is a kind of incremental pathological process. In the last50years, there has been a large number of relevant research about cause of BPH. Androgen and its receptor play an important role in occurrence and development of BPH. But it is not the only cause of BPH. It is not a theory to be able to determine the causality. Although it has been found immune cells infiltration and inflammation in existence in tissue sample of BPH, but this phenomenon was ignored by most clinical and pathological doctor, underestimated the inflammatory process play a key role in occurrence and development of BPH. Inflammatory immune process causes and symptoms of BPH progress may play a key role. The pathological and histological study found that prostate immunology change usually ahead of the histologic BPH. In the development of BPH, immune inflammation and stromal cells function and influence each other, make BPH long-standing in chronic inflammatory immune response process. Prostate immunology has become a new field of study in cause of BPH, people began to research inflammatory and BPH. The cause and characteristics of the increase of white blood cells in BPH. In the process of immune with BPH, some reserches focus on BPH organization cells, immune cells, promote inflammation cell factors in the regulating effect and biological function inflammatory reaction process in prostate molecules immune network. Sex hormones and immune inflammation in occurrence, development of BPH has become a hot issue of prostate immunology. Histological prostatitis (histological prostatitis) cause is not yet clear. The main limitation of tissue samples for difficulty lies in the diagnostic criteria of unity and not, lead to its research has not won its development, physiological function of prostate tissue, the influence of the prostate with all kinds of clinical symptoms occur correlations also lack of further research. So it is necessary to histological prostatitis etiology and its influence of the prostate symptoms of further research. At present the specimen obtain histological prostatitis mainly from patients with prostate hyperplasia, so the histologic prostatitis research usually pay attention to the prostatic hyperplasia.Vitamin D is1,25-(OH)2D3, it plays a part in the body through the vitamin D receptor (VDR). VDR is one member of the steroid hormone/thyroid hormone receptor superfamily, which is the nuclear macromolecules mediated1,25-(OH)2D3play the biological effects. VDR presents in the cell nucleus, it is one member of nuclear receptor superfamily. Human VDR gene is located in12q13-14, stretches approximately78000bp, by11explicit the son, and11of introns. VDR divided into nuclear receptor (nVDR) and membrane receptor (mVDR)2class. NVDR affects gene expression, control the corresponding protein synthesis, mVDR is mainly involved in the maintenance of calcium phosphate balance. The human body cells are almost all the existence of VDR.1,25-(OH)2D3has a variety of biological function in the body, such as regulation immune response, control cell proliferation and differentiation, body mineral balance, its main functions are mediated by VDR.VDR has extensive biological effects, including in maintaining the stability of the serum calcium and phosphorus, regulating cell proliferation, differentiation and immune adjustment function, etc. Its and1,25-(OH)2D3combination can change VDR transcription efficiency. Regulating calcium phosphate metabolism is the main function of VDR. In recent years, it was be found that it also have been an important regulatory role in immune function. VDR and a variety of the happening of the disease have relevance, has become the hot spot of the medical research in recent years. Single nucleotide polymorphism (SNP) is the most common in the human genome, the most widespread DNA polymorphisms type. It has a SNP in every1000bp. It is the human genome physical maps of ideal genetic markers, can meet the metabolism, growth and the orientation of disease genes. It is be considered to be the third generation of genetic markers after the restriction fragment length polymorphism and microsatellite polymorphism. There are about3million SNP in the human genome. The frequency of the SNP is not less than1%in population. SNP showed polymorphism involves only to a single base of variations that can be made of a single base conversion (transition), namely a purine replaced by another purine, or a pyrimidine into another replace a pyrimidine; or transversion, namely purine and pyrimidine exchange between the site. It also a result of insert or missing of the base. The proportion of transtion and transversion is2:1. SNP there are several characteristics:(1) high density. SNP is by far the most widespread in the human genome, the most number of DNA polymorphisms type, about90%of the human genetic variation is the SNP. And in the whole genome STR about15KB have a STR site.(2) short clips. The SNP loci blocks more short, easier for PCR amplification, and the length of the product is less than100bp, this and300～400bp than STRs can better apply to degradation DNA samples.(3) the genetic stability strong. STR in existing in the genome instability and density is lower, the SNP has higher genetic stability, and can be used in the study of population genetics.(4) easy to high throughput, automatic analysis. Most of the performance for the second class SNP a gene markers and the test only need through the simple "+/-" genotyped, easy for classification and determine the gene frequency, be helpful for the laboratory to the standardization of gene type and quality control, and is suitable for fast, high throughput detection and automatic analysis.In theory, the SNP are generally considered to be a bi-allelic markers. In the genome SNP screening often needs only (+/-)analysis. Now SNP is one focus of the human genome research. It has inestimable influence in common diseases in the genetic susceptibility, disease prevention, diagnosis and treatment for cancer.Existing research results show that different people in the genes exist VDR SNP differences. Due to the differences of SNP race there, leading to different ethnic groups for some disease has a genetic easy infectious.There is f/F single nucleotide polymorphisms in VDR gene promoter Fok I site. Fok I site polymorphism caused the differences of VDR mRNA in the level transcription and expression. It cause the different of VDR activity. The change of VDR activity can lead to many of its physiological function change. The histological prostatitis have relationship for one of the functions is immune regulation.At present, have been found on the genes of the existence of VDR and disease of the genetic polymorphism Bsm I, Taq I, Fok I were cut site, which is located in the Fok I promoter, start to change because of the substructure of expression will be produced important regulatory role. Of the existing research found that VDR genetic polymorphism with some chronic inflammatory the happening of the disease have particular concern. WangZhenHua found that the gene polymorphism VDR distribution difference and pediatric repeatedly has certain correlation respiratory infections. They selected is Bsm I polymorphism site, respiratory infection of children repeatedly found B allele frequency have higher trend. ZhangLi such as of124cases of chronic periodontitis patients and91cases of healthy controls were analysis found that, on Taq I sites of VDR polymorphism, t allele can improve the han people in China with periodontitis praecox the risk. Dr such as using Meta analysis to Taq I with chronic periodontitis susceptibility gene polymorphism, found that the388chronic periodontitis patients and355controls were Taq I with chronic periodontitis the susceptibility of business. Therefore, VDR genetic polymorphism and noninfectious inflammatory disease relationship deserves further study.In the analysis and research of many of the SNP method, through the polymerase chain reaction-restrictive fragment length polymorphism means (PCR-RFLP) for now uses more mature and very method. The basic principle is:first use DNA extraction separation and purification purpose gene fragments, and then use PCR technology on purpose gene fragments specificity amplification, and then cut for special enzymes sites on purpose gene fragments of biological enzyme cut, finally to cut the enzyme after fragments electrophoresis, reading their genotype. One PCR technology after years of development, has become a mature genomics of experimental platform, for purpose gene amplification. And also in progress RFLP technology and development for the SNP related research provides a powerful technical support.Objective:The purpose of this study is analysis the relationship VDR gene promoter Fok I sites SNP with BPH complicated histologic prostatitis by researching VDR gene promoter F Explore VDR gene promoter are Fok I sites with histological prostatitis SNP happened between the risk, Fok I SNP genotype. Discusses whether the histologic prostatitis happened genetic predisposition.Through the analysis of VDR gene promoter are Fok I SNP and histological prostatitis locus of points, the relationship between the guidance for histologic prostatitis pathological points and the clinical treatment of guidance to provide certain reference.It is explored the heredity factors in the cause of histologic prostatitis by this artical. The result may provide some clues in further studying development process of BPH.Methods:Collecting August2008to November in Guangzhou Red Cross hospital uropoiesis surgical treatment of prostate hyperplasia merger histologic prostatitis patients169cases (inflammation group), postoperative histopathologic examination reealed prostate hyperplasia, chronic prostatitis, mean age73.9±2.7years old. At the same time data of156patients with simple prostatic hyperplasia (control group), pathological examination reealed prostate hyperplasia, mean age73.56±2.68years old. Two groups of patients are through the urethra prostate specimen cutting (TURP) postoperative gain. After get specimens, the conventional line of10%in darfur-marin soaked. Do conventional colonoscopy. Two groups of patients, and collect relevant basic clinical material, PSA, IPSS score, prostate gland volume, urine flow rate, prostatic hyperplasia drug treatment, etc.VDR gene promoter Fok I SNP genotype are protected by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). The diagnosis of BPH or BPH complicated histologic prostatitis by conventional pathological methods. The relationship between risk of VDR gene promoter Fok I sites SNP with BPH complicated histologic prostatitis was analysed by Logistic regression model. The difference of two groups were analysed by statistical methods.1. The gene bank from NCBI in the relevant information retrieval VDR genes, and obtain the genetic sequence,2. Genefisher software design using PCR primer,3. Use TIANamp Genomic DNA Kit Kit extraction purpose gene segments of DNA,4. In the PCR on the instrument purpose gene amplification,5. Identify the purpose of pieces after amplification fund6. After the expansion of purpose gene fragments specificity enzyme cut,7. Collect enzyme after cutting the fragments agarose gel electrophoresis,8. The results of electrophoretic interpretation. Using conventional pathological methods to check and pure BPH BPH merger histologic prostatitis of occurrence, and on the basis of the research object group.The statistical treatment:All data using SPSS13.0statistical software analysis. Benign prostatic hyperplasia histological prostatitis and simplicity with prostate hyperplasia of the patient’s age, PSA, IPSS score, prostate gland volume, urine flow rate, and other relevant clinical data comparison inspection by t. According to the Hardy-weinberg’s law, adopt direct calculation method for calculation of two groups of genetic type and allele frequency of the actual number and the expected number, understand each type gene in the crowd of genetic balance with degree, the χ2inspection. Compare two sets of genetic type and allele frequency differences, using χ2inspection. Comparing different degree of histological prostatitis genotype and allele frequency differences, using x2inspection. The single factor Logistic regression analysis VDR gene promoter are Fok I SNP different genes with BPH histologic type with risk of prostate inflammation of the correlation between. P<0.05instructions are statistically significant differences, P<0.01that there is a significant difference of statistical significance.The research content and process:The study was made by control studies. Two groups of subjects were divided by into and exclusion standard.The DNA of169patients with BPH complicated histologic prostatitis and156with BPH were abstracted from blood respectively.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used in determining VDR genotypes.BPH and BPH complicated histologic prostatitis were diagnosed by using conventional pathology.The differences of two groups object basic clinical data were analysed by statistical software.The object group representative and Hardy-weinberg balance inspection were calculated by the direct calculation method and statistics method.The relationship of VDR gene promoter Fok I sites SNP with BPH complicated histologic prostatitis was analysed by statistical methods.Results:For benign prostatic hyperplasia histological prostatitis and merger pure prostatic hyperplasia VDR gene Fok I sites with SNP genotype distribution of the actual number and the expected number directly computing, again through the χ2inspection, the results show the actual number and the expected number was not statistically significant difference (P>0.05), meet Hardy-Weinberg’s law, in this population genetics explain representative data.In two groups of patients with relevant clinical data of comparison with t test. The results suggest that both groups of patients are representative data.In two groups of patients with relevant clinical data of comparison with t test. The results suggest that both groups of patients, average age no significant difference (P=0.118), inflammation group PSA levels (ng/ml) significantly higher than those in the control group (P=0.001). Two groups of patients preoperative routine blood and urine check has not seen the infection, liver, kidney check has not seen the exception. International prostate symptoms (I-PSS) score:inflammation for15.4±2.5points group, compared with15.3±2.32points, the difference between the two groups was statistically significant (P=0.744). Two groups of patients through B ultrasonic measure before the abdomen prostate olume, and use around, front and back, and three groups, diameter line (mm) said, inflammation group prostate olume (ml)=0.52×(prostate is the product of the three diameter) significant higher than control group (P=0.027), postoperative prostate removed weight (g) was not statistically significant (P=0.423) inflammation group has35patients preoperative no acute urinary retention, routine urine flow rate do check, the control group40routine urine flow rate (ml/s) check, two groups of the rest of the patients were already stay urinary catheter, between the two groups have done urine flow rate of the difference was not statistically significant (P=0.626).Two groups of patients for VDR gene promoter Fok Ⅰ SNP compared gene loci type, the χ2inspection. Results showed that genetic type of inflammation in ff was37%(62/169), the control group was25%(39/156), the genetic type in the distribution difference between the two groups was statistically significant (P<0.05), and ff and ff genotype between the two groups for:the distribution of inflammation group:25%(43/169); The control group:35%(55/156) and38%(64/169);40%(62/156), the difference was not statistically significant meaning (P>0.05). F gene frequency in inflammation group is44%(150/338), the control group was55%(172/312), F gene frequency in inflammation group is56%(188/338), the control group for:45%(140/312), the difference between the two groups was statistically significant (P<0.05).The research selected the crowd moderate to severe histologic prostatitis three genotype distribution is not in conformity with the Hardy-Weinberg’s law, so this type in this study does not have the crowd representative. And mild and moderate histologic prostatitis three genotype distribution meet Hardy-Weinberg’s law. It analyzes three genotype in mild to moderate histologic prostatitis differences between the. Results showed that FF, FF and genotype mild inflammation in FF group and moderate inflammation of distribution between for:32%(25/79),23%(12/52);46%(36/79),40%(21/52);22%(18/79),37%(19/52), the difference was not statistically significant meaning (P>0.05). F gene frequency in mild inflammation group and moderate inflammatory group were54%(86/158),43%(45/104), F gene frequency in both groups were46%(72/158),57%(59/104), the difference between the two groups was statistically significant (P>0.05).Through the gene sequencing method to the experimental result of the PCR-RFLP analysis, three kinds of random genetic type,10cases of the gene sequencing, discovered genes with before the experiment of gene content. Verify the accuracy of PCR-RFLP experiment and repeatability.To VDR gene promoter Fok I site F/f SNP genotype as the independent variable and merger with BPH histological prostatitis as the dependent variable, the merger BPH histological prostatitis risk factors Logistic regression analysis. The results showed that in the control factors such as age, Fok I polymorphism is prostate hyperplasia merger histologic prostatitis happen risk factors (P<0.05). One genotype relative to ff ff genotype OR value of2.03(0.279～0.866) which ff genotype relative to ff genotype OR value of1.32(0.446～1.287), including ff genotype relative to ff genotype OR value of1.54(0.382～1.105).Conclusion:1. VDR gene promoter Fok I sites in genotype and the SNP ff gene allele frequency in f prostate hyperplasia histological prostatitis and pure with the distribution of patients with prostate hyperplasia difference was statistically significant, explain ff genotype and histological prostatitis occurs with correlation. VDR gene promoter Fok I SNP sites as a genetic markers that histologic prostatitis occurs with the crowd of genetic predisposition.2. Through the single factor Logistic regression analysis VDR gene promoter Fok I sites and prostatic hyperplasia with SNP histological prostatitis happen, the relationship between the found VDR gene promoter Fok I SNP sites is histologically prostatitis happened independent risk factor. From the genetic level for treatment of prostatitis histologically to provide the reference.3. VDR gene promoter Fok I sites different genotypes of SNP the distribution and histological prostatitis serious degree of difference was statistically significant, need to undertake more research reveals the sample SNP sites and histological prostatitis pathological points the correlation of type, thus for pathology classification and clinical treatments with certain guidance.4. Histologic prostatitis affect the PSA and prostate voleume to a certain extent. |
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