| Background:The gallbladder is an important part of biliary system,it has the function of stores bile, concentrated bile, secrete mucus and emptying function. At the same time also have played an important role in coordinating biliary pressure. The gallbladder contraction function by neuromodulation and humoral regulation,however the humoral regulation of gallbladder movement regulate the dominant position. The gallbladder contraction element (CCK) is secreted directly involved in the control of the effect of biliary movement is especially important, and is considered to be the main way(1). In the process of gallbladder contraction autonomy, gallbladder of spontaneous excitability in the heart, the same with spontaneous gastrointestinal tract has the same relationship between excitatory organs and characteristics,and it can be inked to these specific differentiation of cells.In 2007, Hinescu ME(2)asseted the effect for the first time on the disease-free gall bladder resection in patients with certain ICCs (interstitial cells of cajal, cajal interstitial cells),which have specific c-kit tyrosine kinase receptor. The ICC cells is a kind of special mesenchymal cells have a variety of functions, through the experimental research in recent years, most scholars believe that biliary ICCs sample cells and gallbladder can produce and disseminate spontaneous rhythms, regulate gallbladder movement, cause the gallbladder contraction function such as closely related(3,4).At present the research of gastrointestinal ICC has been very deep, is the initial cause of gallbladder emptying dysfunction and the key dynamic factors’6’. IT has confirmed that the ICC is involved in regulating gastrointestinal tract neurotransmitters process, which is the pacemaker of gastrointestinal smooth muscle movement and passing factor(7). This class cells and gastrointestinal ICC have similar morphological characteristics and physiological functions, but many studies have also shown it has obvious difference with the gastrointestinal tract. The structure of gallbladder wall and gastric wall is similar so as to the ICC in the gastrointestinal tract and whether the ICC distribution and morphology of the gallbladder are similar too? And whether at the same time corresponding reflected in gallbladder movement mechanism also plays a similar control effect? These questions guided us from cellular level to analyze the ICC cells in the biliary system, by the ICC and the gallbladder to study the dynamic changes of the relationship between the point of view.In most of the gastrointestinal tract disease, most of the occurrence of diseases is always accompanied by the ICC to reduce or missing,also has been linked with exercise correlation disease in the first place, such as constipation, diabetic gastroparesis(5’8), in the biliary system is one of the most common cholelith disease. Cholelith disease has its pathogenesis in addition to "supersaturated" cholesterol in bile and cholesterol "micro crystal nucleation time shorten" theory, gallbladder’s own factors (gallbladder motor function impairment) role is very important,too(9).The treatment of diseases, surgical treatment mainly include open cholecystectomy, laparoscopic cholecystectomy and gallbladder stone extraction, but in view of the operation risk, therefore, in recent years through the way of combining traditional Chinese and western medicine treatment of the disease have achieved ideal curative effect. Chronic calculous cholecystitis in TCM belongs to the category of "up" and "jaundice", belongs to liver and gallbladder damp and hot card on pathogenesis, therapeutic principle for heat dehumidification, liver and gallbladder, rows of stone. Bian Yang Ming artemisia capillaris soup originated from zhang zhongjing "treatise on febrile diseases", which was pulse license and cure. Herba artemisiae capillaris, gardenia, rhubarb na are in this old formulas. The formulas in the party for the treatment of Yang huang classic, but proved,without jaundice and have courage certificate of damp and hot, with the effect of even more significant. Our study shows that using the artemisia capillaris soup to chronic calculous cholecystitis have obvious relief, the formulas for disease produces in the process of dynamic change of the link between the ICC cells and the relationship between chronic calculous cholecystitis, subject to our further research.Objective:Section 1:Application of artemisia capillaris soup for treating chronic calculous cholecystitis patients curative effect and the influence of the gallbladder contraction function analysis.Section 2:By feeding high cholesterol stone to feed 8 w model of chronic calculous cholecystitis, and the treatment group given under the condition of the high cholesterol feed 8 w at the same time lavage artemisia capillaris soup. Each group for HE staining and transmission electron microscopy (sem), muscle, the muscle tone and used in the determination of calcium ions in the cells.1. Observe the normal group, the high cholesterol group and treatment group after HE staining showed the change of the inflammation of the situation, and the ICC in three groups under tem cell ultrastructure change.2. Observe the three group of gallbladder muscle strips in the gallbladder contraction, gallbladder muscle strips of muscle tone changes.3. Observe the original generation after cultured Cajal interstitial cells, light microscope and fluorescence microscope observation.Methods:Section 1:1. To collect the first affiliated hospital of dalian medical university in January 2010 to December 2011 specimens of outpatient and inpatient, according to the diagnostic standard and discharge index of specimens were selected for the number of cases, determine the cases into the group. At the same time choose no associated biliary diseases volunteers as control group.1.1 Treatment group:28 cases, male 18 cases,10 cases of women, aged 35 to 60 years, an average of 50.9+/-20.1 years old.1.2 In the control group:30 cases,21 cases of male, female 9 cases, aged 25 to 60 years, an average of 52.7+/-18.8 years old.2. Two groups of patients after outpatient departments or anti-infection and correct electrolyte disorders after admission to hospital after treatment, the control group given oral placebo (herbal tea), the treatment group oral artemisia capillaris soup. The clinical manifestations of the two groups was observed, and the curative effect evaluation.3. Statistical control and tested before and after treatment in patients with ultrasound were used to detect the gallbladder contraction function approach to measure the fasting state of gallbladder volume, eating fat postprandial gallbladder volume and gallbladder contraction ratio, and the data also entry in the database for statistical analysis.4. Before and after treatment in patients with the control group and tested for the statistical analysis of the gallbladder contraction ratio observed patients and patients with a control index of the differences and changes of these indexes before and after treatment.Section 2:1. Choose healthy test o-level guinea pigs, all male, were randomly divided into three groups:normal group, model group and treatment group. Feeding standard guinea pigs feed normal group, model group and treatment group was given compound good cholesterol stone to feed, for eight weeks,after the treatment group uses the artemisia capillaris soup for lavage. Normal group and model group with the same volume of physiological saline lavage, and breeding programs before you continue, a total of 4 weeks.2. Tissue of each guinea pig gallbladder was taken. Through steps such as embedding, sectioning and staining, we stained it with HE. We observed pathological changes of gall bladder in each group under light microscope. Let pathology teacher assess the degree of gallbladder inflammation to apply the independent double-blind principle. So we could verify whether the establishment of gallbladder model was successful.3. Take each gallbladder tissue, by double fixed, gradient ethanol dehydration, embedding, sectioning and staining steps, such as the ICC cell ultrastructure observed under transmission electron microscope.4. Take each gallbladder tissue, article made the gallbladder muscle tension transducer, under the stimulus of gallbladder contraction, and through multi-channel biological signal collection and analysis system, record of three groups of gallbladder muscle contraction activities frequency and amplitude.5. Take each gallbladder tissue cut into blocks, sterile conditions by collagenase digestion method into cell suspension inoculation to medium after 3-5 d, going to do the immunofluorescence stainingResults:Section 1:1.Clinical treatment group total effectiveness 75%, control group total effective rate was 12.1%, and the treatment group was better than control group (P< 0.05). Results indicate that in artemisia capillaris soup for treating chronic calculous cholecystitis with better clinical curative effect, and has good application value.2.1 Before the treatment of patients with chronic calculous cholecystitis gallbladder contraction ratio than the control group the gallbladder contraction ratio decreased significantly (P< 0.05), its function of gallbladder contraction performance in bad condition.2.2 Taking the artemisia capillaris soup treatment, tested the gallbladder contraction function in patients with chronic calculous cholecystitis was significantly increased (P< 0.05).Section 2:1. HE staining under light microscopy observation three groups of inflammatory degree has obvious difference; ICC cell ultrastructure under electron microscopy (sem) and with the surrounding smooth muscle cells, the structure of the gap junctions between nerve cells have obvious difference.2. The gallbladder muscle experiments, the amplitude and frequency of model group compared with normal group had significantly lower (P< 0.05), the treatment group on the amplitude and frequency of relative model group significantly had increased (P< 0.05).3.Stable in vitro cultivation gallbladder ICC cells, and through the c-kit immunofluorescence staining appraisal under the condition of in vitro culture has a typical long shuttle type of cells to the ICC.Conclusions:1. A significant reduction in the gallbladder contraction function in patients with chronic calculous cholecystitis, after artemisia capillaris soup gallbladder contraction function increased significantly after treatment.2. In the process of chronic calculous cholecystitis formation, the ICC cellular structure change gradually, and with the surrounding of smooth muscle, nerve cell connections also change accordingly. |
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