| The earliest record of human for hemorrhoids in770BC, but to this day, hemorrhoids is what?What is the nature of hemorrhoids?Within the academic circle is still in aimless and endless debate, we have to face is that our main pathogenesis is not fully determine for hemorrhoids. Along with the science of advancing with The Times, some scholars constantly committed to the discussion and the research of hemorrhoids, through hard work and effective research and discussion, until the70s, we have put forward the landmark "anal cushion down theory", and then further enriched and developed in the study of the theory.Although hemorrhoid "anal cushion theory" is acknowledged by most scholars, but according to current research foundation, still can’t explain the exact nature of the hemorrhoids, lead to pathological changes of anal cushion is what? So far the research theory is not targeted to answer to this problem., therefore, the current research on "theory of anal cushion down" is just at the initial theory to explore stage, hemorrhoids tissue pathological changes of molecular biology research is lack of effective clinical and experimental proof, also the characteristics of anal cushion can not truly guide clinical application.This thesis in yiqi lifting method in treatment of prolapse hemorrhoids achieve good clinical curative effect, on the basis of application of molecular biology research means, determination of the degree of prolapse of hemorrhoids specimens of Fibulin-3, Fibulin5mRNA and protein expression, by studying the different degree of hemorrhoids prolapse sex organization Fibulin family mRNA and protein expression level difference, investigate Fibulin proteins family internal connection and prolapse of hemorrhoids occurrence and development.At the same time, this study applies the means of clinical analysis and molecular biology research, yiqi granules in the repair after the intervention, analyzing the internal organization of mRNA expression of dynamic Fibulin proteins family table, and the further study of yiqi lifting method in treatment of prolapse of hemorrhoids mechanism, reveal yiqi lifting method by slowing down aging Fibulin proteins family, in treatment of prolapse of hemorrhoids disease pathways. Design the starting point of this experiment is to seek evidence of molecular biology for anal cushion down theory; For the theory of traditional Chinese medicine diagnosis and treatment of prolapse hemorrhoids with modern scientific basis.ObjectiveThis study aims to through the following two ways:(1) Way to varying degrees, and to examine the sexual hemorrhoids prolapse organization Fibulin anti-aging proteins in the family (note:Fibulin-3and Fibulin-5) mRNA and protein expression level;(2) The way in yiqi lifting method after the intervention, the cases of hemorrhoids II degrees Fibulin proteins family mRNA expression in organizational change.This study hopes to study two purposes:(1) Purpose Fibulin proteins family and sexual hemorrhoids prolapse, and to examine the occurrence, development whether there is a correlation?If there is a correlation, so how do exist correlation?(2) The purpose, the further study of yiqi lifting method in treatment of prolapse of hemorrhoids mechanism:whether yiqi lifting method by slowing down aging Fibulin proteins family, play a role in treatment of prolapse of hemorrhoids?To further elucidate the pathogenesis of hemorrhoids prolapse sex breach, at the same time for treatment and prevention of the disease mechanism.MethodsExperimental (A)1. Grouping schemeThe object of study from2010January to2013December in Guangzhou University of Chinese Medicine First Affiliated Hospital anorectal Inpatient and outpatient patients, a total of64cases. The grade Ⅱ in21patients, grade Ⅲ21cases,22cases of fourth degree hemorrhoids patients. At the same time,22cases of low simple anal fistula patients, as control group.2. Specimen collection and extractionIn the second, third degree, fourth degree haemorrhoids patients receiving "hemorrhoids resection", take resection hemorrhoids tissue as the specimens; low simple anal fistula patients."The low anal fistula resection" in the process, there were no changes from "anal cushions" normal tissues as contrast samples. The experimental group and the control group of specimen length were0.5cm,30min in vitro under aseptic condition immediately as backup save-80℃refrigerator or liquid nitrogen.3. The experimental method①Total RNA extraction and reverse transcription②The real-time quantitative PCR detection of Fibulin-3mRNA level③The detection of Fibulin-3protein expression, Western blot4. Statistical processingUsing SPSS13.0software process the data. The data to mean+/-standard deviation (X+S) said. Each Fibulin-3expression results between independent sample t-test and single factor variance analysis.:difference was statistically significant (P<0.05); There are significant difference (P <0.01) for statistical significanceExperimental (A)1. Grouping schemeThe object of study from from January2010to December2010, guangzhou university of Chinese medicine the first affiliated hospital anus bowel division. The inpatient and outpatient patients, a total of62cases.The Ⅱ hemorrhoids patients included in20cases, Ⅲ degrees in20cases, Ⅳ hemorrhoids hemorrhoids patients included in22cases. At the same time, choose low simple anal fistula disease in17patients, as control group Collection2. Specimen collection and extractionIn Ⅱ degrees, Ⅲ degree, Ⅳ hemorrhoids patients received, after the "hemorrhoids resection" return resection hemorrhoids organization as the specimen; Low in simple anal fistula patients during the process of "low" anal fistula resection, extraction of patients, there was no pathological changes of "anal cushion" normal tissue specimens for comparison. The specimens of the experimental group and control group based on length are0.5cm, in vitro aseptic conditions within30minutes immediately as to-80℃refrigerator or kept in liquid nitrogen. Set aside.3. The experimental method①Total RNA extraction and reverse transcription②The real-time quantitative PCR detection of Fibulin-5mRNA level③The detection of Fibulin-5protein expression, Western blot4. Statistical processingUsing SPSS13.0software process the data. The data to mean+/-standard deviation (X+S) said. Each Fibulin-5expression results between independent sample t-test and single factor variance analysis.:difference was statistically significant (P<0.05); There are significant difference (P <0.01) for statistical significance.Experiment (C)1. The grouping schemeThe object of study from2010-2013in the first affiliated hospital of guangzhou university of TCM anorectal hospital wards and Clinic patients, a total of49cases. All patients were diagnosed as II degree of hemorrhoids. Patients were randomly divided into four groups (treatment group3groups; control group1).Among them, the Ⅱ degrees hemorrhoids treatment group (group A):1n=11cases; Ⅱ degree of hemorrhoids treatment group (group B),2n=12cases; Ⅱ degree of hemorrhoids treatment group (group C)3, n=9. Group with the control group1:n=7).2. Specimen collection and extractionEach intervention method is as follows. Group A:taking fill in beneficial gas particles; Group B:oral sweet clover flow immersion; Group C:taking fill in beneficial gas particle flow immersion+sweet clover. Only as a blank control group D.2w as one period of treatment, all cases ended after2courses of treatment.2days after treatment has stopped, the patient clinical observation and data analysis.Arrange local infiltration anesthesia in patients with downward "hemorrhoids resection, and for the cases of postoperative file. usd10.00Fibulin hemorrhoids samples-3mRNA, Fibulin5mRNA expression of testing.3. The experimental method①Detection of safety indicators:General physical examination project, blood, urine, stool routine, liver and kidney function examination, ecg event record check;②Detect curative effect index1:The change of clinical manifestation③Detect curative effect index2:Testing each sample in the organization Fibulin-3mRNA expression level; Testing each sample in the organization Fibulin5mRNA expression level.4. Statistical processingCount data with X2test, percentage level data and comparison with Ridit analysis, measurement data using analysis of variance and t test.Using SPSS10.0software for analysisResultsResults (1)1.Each sample object Fibulin-3mRNA expression levels were compared, including grade II, III and IV grade grade,"internal" and "control group" operation specimens can be detected the expression of Fibulin-3mRNA (Fibulin-3mRNA=0)."Fibulin-3mRNA expression was0.68±0.37hemorrhoids group" specimens "control group" specimens, Fibulin-3expression level of mRNA was0.96±0.43, found that:"in a relatively low level of Fibulin-3mRNA expression by the group" samples, through independent samples t test, the difference between the two groups has statistical significance.2. In the "internal hemorrhoid group":expression of the expression of Fibulin-3mRNA grade group was0.69±0.17, Ⅲ grade group of Fibulin-3mRNA was0.56±0.15, fourth degree hemorrhoids group Fibulin-3is0.42±0.22mRNA. Comparison between the groups, with the degree of clinical prolapse increased Fibulin-3mRNA expression decreased gradually, and the comparison of the two two groups showed, each between two groups (between Ⅱ grade Ⅲ grade between group and group Ⅲ grade group; group Ⅱ and Ⅳ grade grade group and the fourth degree haemorrhoids group contrast group), the differences were statistically significant.3. The expression of Fibulin-3protein in the samples of each level are compared,"internal" and "control group" operation specimens have the expression of Fibulin-3protein."The control expression of protein Fibulin-3group" specimens was1.15±0.53;"hemorrhoids resection specimen group" Fibulin-3protein expression was0.77±0.47."The expression of internal hemorrhoid group" of the Fibulin-3protein in the lower level, the independent samples t test, a statistically significant difference between the two groups.4. Expression of Fibulin-3protein in different "hemorrhoids group" is different:①Ⅱ Fibulin-3protein expression in the grade group was0.79±0.35; grade Ⅱ Ⅲ Fibulin-3protein expression in the group was0.57±0.37;③grade IV Fibulin-3in the group of RNA protein expression was0.36±0.29. Among the two two comparison results suggest that, between the two groups (each group Ⅲ grade Ⅱ and grade Ⅲ grade group; group and the fourth degree haemorrhoids group; second grade and fourth degree haemorrhoids group contrast group), the differences were statistically significant.Results (2)1. Comparing the expression level of Fibulin-5in samples of each research group mRNA:Grade Ⅱ, Ⅲ and Ⅳ grade grade,"internal" and "control group" operation specimens were detected the expression of Fibulin-5mRNA. Among them,"Fibulin-5mRNA expression was0.32±0.22hemorrhoids group" specimens "control group" specimens, Fibulin-5expression level of mRNA was0.83±0.31,"Fibulin-5mRNA expression by the group" samples is low, the independent samples t test, a statistically significant difference between the two groups.2. The "internal" group, expression of different groups of Fibulin-5mRNA is also a clear distinction:the expression of Fibulin-5II grade group mRNA is0.52±0.27; grade Ⅱ Ⅲ Fibulin-5expression in mRNA group was0.37±0.12; expressions of Fibulin-5grade IV in group mRNA was0.23±0.20. Among the two two comparison result also shows that, between the two groups (each group Ⅲ grade Ⅱ and grade Ⅲ grade group; group and the fourth degree haemorrhoids group; second grade and fourth degree haemorrhoids group contrast group), the differences were statistically significant.3. The expression of Fibulin-5protein in the samples were compared, results showed that:"internal hemorrhoid group" and "control group" operation specimens have the expression of Fibulin-5protein. Among them,"control the expression level of Fibulin-5protein group" specimens was1.10±0.47;"by the group" the expression of Fibulin-5protein expression was0.58±0.45,"internal" Fibulin-5protein showed low expression level, the independent sample t test.Inspection, statistically significant difference between the two groups.4. In the different "hemorrhoids group", the expression of Fibulin-5protein is different:①Fibulin-5grade Ⅱ group expression was0.79±0.33; grade Ⅱ Ⅲ Fibulin-5protein expression in the group was0.61±0.21; protein Fibulin-5and Ⅳ grade group expression was0.44±0.26. Each of the two groups of comparison, the differences were statistically significant.Results (3)1. Application of the different drugs clinical efficacy and prognosis of stem the "treatment group" and "the blank control group","A"(Buzhong Yiqi granules) efficiency is81.8%,"B group"(wood Xi extract tablet) efficiency is75%,"C group"(Buzhong Yiqi granule+clover tidewater dip liquid film) efficiency is100%. The1group,2group,3group as the object of observation, calculation of second degree hemorrhoids treatment group (total) efficiency is84.4%, higher than the "D" efficiency is23.33%, the effective rate of treatment in the two groups were statistically significant differences in contrast,(P<0.01)2. The second degree hemorrhoids treatment group and the control group tissues are available to the Fibulin-3stability of mRNA:"Fibulin-3mRNA expression was0.74±0.18in group A were" Fibulin-3"; the expression level of mRNA was0.68±0.24in group B were" Fibulin-3"; the expression level of mRNA was0.88±0.14the C group" Fibulin-3"samples, the expression level of mRNA was0.53±0.17in group D were" hemorrhoids. In addition,"D" and "A group","B","C group" two two "comparison, shows low expression of Fibulin-3mRNA expression of D" group organization:"D group" in the sample of Fibulin-3mRNA expression and "A","B","C group" Fibulin-3mRNA expression in specimens with different (independent samples t test), the difference was significant (P<0.05)3. The second degree hemorrhoids treatment group and the control group were able to detect Fibulin-3mRNA, including:"Fibulin-5mRNA expression was0.50±0.14in group A were "Fibulin-5", the expression level of mRNA was0.47±0.15in group B were "Fibulin-5"; the expression level of mRNA was0.65±0.19C group" specimens,"Fibulin-5mRNA expression was0.44±0.11D group" specimens. In addition,"D" and "A group","B","C group" two two are compared, results show that the "D group" Fibulin-5mRNA "in the expression of low expression state":"and each treatment group showed Fibulin-5mRNA expression were differentially expressed in D group" in the sample of Fibulin-5mRNA (the independent sample t test), the difference was significant (P <0.05) ConclusionConclusion (A)Deletion of Fibulin-3may lead to increased MMPs activity, and MMPs activity increased is the speed of elastic fibers, elastic force local tissue disintegration and promote decline, one of the important reasons for the occurrence of prolapsed internal hemorrhoids.Conclusion (B)Deletion of Fibulin-5may lead to impairments in elastic protein synthesis, and elastic fiber synthesis is disorder resulting elastic fibrous lesions, pelvic floor structure relaxation, supporting the role of local tissue elastic force decreased, and so one of the important reasons of prolapsed internal hemorrhoids.Conclusion (C)Different interventions include Qi lifting method, is adopted to improve the Fibulin-3mRNA (or Fibulin-5mRNA) expression and ultimately play a therapeutic role.Conclusion (D)To improve the Fibulin-3mRNA (or Fibulin-5mRNA) expression may be a means of treatment of second degree hemorrhoids prolapse performance..Conclusion (E)Different treatment to improve the Fibulin-3mRNA (or Fibulin-5mRNA) of the differences of expression.Conclusion (F)Second degree hemorrhoids prolapse symptoms as the main performance of the self-healing certain... |
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