Discussion About Prevention And Treatment Of Liver Cancer Experience Connotation Of Professor Sun Gui-zhi From Anti Liver Fibrosis Effect Of "Taohong Qizhu Ruangan Decoction"

Gui-zhi sun is a famous professor of Chinese and western medicine co mbined treating cancer, she has engaged in clinical work of tumor prevent ion and treatment for nearly 60 years, treat of hundreds of thousands of middle-late tumor patients, especially in the treatment of digestive trac t tumor, applying the idea of "cure before ill" to treat liver fibrosi s/cirrhosis, prevention and treatment of liver cancer and has accumulate d rich clinical experience. Mortality of liver cancer lists the third in cancer mortality in China, and liver cancer developing rapidly, relative ly short lifetime. Chronic hepatitis B viral hepatitis is papular in our country, "chronic hepatitis B viral hepatitis-hepatitis liver fibrosis/ cirrhosis -liver cancer" is three stages in the development of liver can cer,studies found that 80% of patients with liver cancer Originated from liver fibrosis, combined with Chinese medicine idea "cure before ill", p rofessor Gui-zhi sun think that for liver cancer prevention shouldbe pri marily prevent, Prevention and treatment shoule be combined, mainlyinclud ing:"prevention before disease" means the prevention and treatment of liver fibrosis/cirrhosis of the liver; "prevent after disease" namely a ctive treatment of liver fibrosis/cirrhosis, prevent progress to liver c ancer; "treat in the early", namely active treatment early liverfibrosi s/cirrhosis of the liver.This subject summarize experience of clinical treatment for liver fi brosis/cirrhosis of professor Gui-zhi sun. selecte clinical cases of prim ary liver cancer with a history of liver fibrosis/cirrhosis to carry out Prospective clinical study,adopts "the scale-free network analysis" tech nology and topic model technology to analyse, carry out in-depth analys is to clinical characteristics andclinical evidence-based medicine law o f the prevention treatment of liver fibrosis and treatment of liver can cer of Gui-zhi sun, on this basis, we took experiments combined inside and outside, combining to the sun old experience in prevention and treatment of liver cancer treatment for liver fibrosis "Taohong qizhu ruangan dec oction" generated from blocking of liver fibrosis, TGF-β/Smad signalin g pathways regulating epithelial mesenchymal into experimental research, from one side to interpret the scientific connotation of academicthought of professor Gui-zhi sun.Main content of this Paper is divided into three parts:The first part:carry out theory analyse of academic thoughts and th eory diagnosis of treating liver fibrosis/cirrhosis and prevention and t reatment of liver cancer of professor Gui-zhi sun.Grasp academic thought of treatment of liver fibrosis/cirrhosis, prev ention and treatment of liver cancer by ancient and modern literature ab out liver fibrosis and clinical study.1. Based on professor Gui-zhi sun theory of "prevention before disea se",treat liver fibrosis from "Yiqi huoxue, ruanjian sanjie" "Buyi piw ei".2. Professor Gui-zhi sun thought of liver cancer treatment shall be based on "Yiqi Huoxue Ruanjian Jiedu" for treatment, commonly use "Huan gqi jianzhong decoction" "Tenghu decoction" "Huoxue lishui decoction", etc.The second part clinical study:to carry out data mining and curativ e effect analysis about diagnosis rule of professor Gui-zhi sun of anti liver fibrosis and prevention and treatment of liver cancer.Analysis clinical information of 35 primary liver cancer cases with a history of liver cirrhosis/liver fibrosis, carry out forward-looking analysis, adopt the scale-free network and topic model technology mining to carry out in-depth study about clinical characteristics, prescription rule of professor Gui-zhi sun.1. The clinical curative effect:patients’objective curative effect of stability factor is higher than before treatment after treatment; Exhaustion, exhausted lazy said god, itchy skin, dry mouth, mouth pain, abdominal distension, pain, irritability, heave a deep sigh, symptoms such as pain, edema improved; ALT、AST、AFP decreased; Albumin increased after treatment; red blood cells, white blood cells, hemoglobin and platelets are higher than before; Quality of life scores have improved obviously.2. Results of data mining:in 35 cases of primary liver cancer patien ts with liver fibrosis/cirrhosis, summed up top four syndromes- "core syndrome",at the same time, according to the drug node, tightness, and frequency of usage, summed up the corresponding four core syndrome of "core prescriptions", among them, the corresponding core "qi and bloo d stasis prescription drugs" for:mountain of vinegar, vinegar turtle sh ell, root of herbaceous peony, rose, green plum, dried tangerine or oran ge peel, pink flower son of hubei, earthworm and so on." "Poison evil in vasion core" prescriptions corresponding drug for:giant knotweed, Chines e gooseberry, spreading hedyotis herb, floor, glabrous greenbrier rhizom e. "Blood stasis in the core formula" corresponding drug for:vinegar tur tle shell, vinegar hill, giant knotweed, pink flower, earthworm, peach k ernel, chicken’s gizzard-membrane, rhizoma zedoariae, etc. "Spleen kidney deficiency core formula" corresponding drug for:radix astragali, radix p aeoniae alba, radix pseudostellariae, fleece-flower root, medlar, fructu s ligustri lucidi, etc. According to the research of the topic model of syndrome with traditional Chinese medicine (TCM) corresponding to higher probability value of 5 groups theme, respectively is:glove two deficie ncy syndrome:multi-purpose pseudostellaria root, angelica, glabrous gre enbrier rhizome, polygala, bran Fried atractylodes drugs; Poison evil in vasion, often with giant knotweed, polyphylla, vinegar added tortoise sh ell and other drugs; Qi and blood depression syndrome is multi-purpose p each kernel, earthworm, vinegar turtle shell, such as the centipede drug s; Blood stasis block, the multi-purpose gun mount armor, vinegar turtle jia drugs;Spleen and kidney deficiency syndrome is multi-purpose astragal us, fleece-flower root,radix paeoniae alba and other drugs; Water stop i ssuing common pixu (spleen deficient) atractylodes, grifola, astragalus r oot and other drugs. According to clinical research and data mining techn ology,the data result and Professor Gui-zhi sun by submitting a paper do cuments, interview investigate, review modification, summed up the Profe ssor Gui-zhi sun experience formulas for the treatment of liver cancer a ssociated with liver fibrosis/cirrhosis, mainly include:raw huangqi, Fr ied atractylodes, radix pseudostellariae, peach kernel, rhizoma zedoariae a nd other drugs. All parties have a disease of the spleen and replenishing qi,promoting blood circulation to remove blood stasis, soft firm, reflect professor Gui-zhi sun teacher righting this culture, yiqi huoxue, soft f irm fights the clinical diagnosis and treatment of thinking.The third part Vitro and vivo experiment validation:discuss the mol ecular mechanism of clinical curative effect of treating hepaticfibrosis, interpret the scientific connotation of professor Gui-zhi sun.In vivo experiment is proposed by intraperitoneal injection of 50% C CL4 olive oil solution Wistar rats preparation of liver fibrosis in rats model, the preparation of liver fibrosis model, building at the same ti me using the "Taohong qizhu ruangan decoction" low, medium and high dos e of intervention therapy, and with colchicine,Fuzheng Huayu recipe. In building for 3,6,9 weeks three time points dynamic observation of bloc king liver fibrosis progression, and TGF-β/Smad signaling pathways regu lating epithelial mesenchymal transformation is directly related to the immunohistochemical protein expression and mRNA expression of indicator s.In vitro experiment select Hep-G2 as low differentiation of hepatic stem cells, in vitro and restructuring TGF-β protein intervention, buil t vitro liver fibrosis model, using "Taohong qizhu ruangan decoction" t o intervention in vitro, use immunofluorescence and Western Blot to test protein expression influenced by "Taohong qizhu ruangan decoction" to H ep G2 cells. Combining the experiments in vivo and in vitro, to explorec urative effect and mechanism of "Taohong qizhu ruangan decoction" treati ng liver fibrosis, to provide experimental basis for interpreting the sci entific connotation of professor Gui-zhi sun.1. The in vivo efficacy analysis:1.1 After drug intervention, different time node degree of liver fibrosi s in rats.3,6,9 weeks of rat liver HE staining and Masson staining prompt:at 3 weeks, part of rat liver tissue happen fibrosis, "Taohong qizhu ruangan decoction" middle dose and high dose group improved the most obvious; at 6 weeks rat liver tissue of Model group form false flocculus, suggesting hepatic fibrosis model built successly, "Taohong qizhu ruangan decoctio n" and Fuzheng Huayu recipe,colchicine have obvious relief for liver fib rosis in rats, "Taohong qizhu ruangan decoction" high dose improved sign ificantly compare to other groups; at 9 weeks rat liver tissue of eachgro up form false flocculus, liver tissue structure of "Taohong qizhuruangand ecoction" high dose group improves the most obvious.Experimental results show that with the extension of building time, e ach dose groups all have the effect of blocking the progress of liver fi brosis in rat models. And campare to the model group, at 9 weeks, effect of "Taohong qizhu ruangan decoction" high-dose group is the most obviou s.1.2 Different timing of rat liver tissue immunohistochemical protein expression results:3,6,9 weeks of rat liver pathological images prompts:immunohistoc hemical study compared with the blank group, model group E-cadherin expr ession decreased in the blank group, and the expression of TGF-β1, Vime ntin and Smad2 enhanced expression in the blank group, indicating that E pithelial-mesenchymal transition continue happens in rat liver tissue,ep ithelium of rat liver fibrosis may be happenes via Epithelial-mesenchym al transition; campared to model control group, "Taohong qizhu ruangandec oction" can increase E-cadherin, reduce TGF-β1, Vimentin and Smad2 prote in expression, indicate that "Taohong qizhu ruangan decoction" by adjust ing the TGF-β/Smad signaling pathways, reduce Smad2 protein expression, s o as to reduce TGF-β1, increase E-cadherin, reduce Vimentin expression, p lay to reverse Epithelial-mesenchymal transition, the effect of treatmen t of liver fibrosis "Taohong qizhu ruangan decoction" middle dosegroup a nd high dose group is the most significant.1.3 The fluorescent quantitative PCR to detect liver tissue E-cadherin, TGF-β1, Vimentin, Smad2 mRNA expression:3,6,9 weeks in the rat liver tissue mRNA expression shows:compare d with the blank group, model group E-cadherin mRNA expression decreased, expression of TGF-β1 mRNA, Smad2 mRNA, Vimentin mRNA increased, promptin the process of the formation of liver fibrosis, ongoing transformationof epithelial mesenchymal of liver tissue of rats, may happened through the TGF-β/Smad signal conditioning; compare with the model control group, "Taohong qizhu ruangan decoction" can increase E-cadherin mRNA, reduce TGF-β1 mRNA、Smad2 mRNAand Vimentin mRNA expression, each dosegroup of "T aohong qizhu ruangan decoction" all have better curative effect, prompt t hat "Taohong qizhu ruangan decoction" could adjust the TGF-β/Smad signa ling pathways to regulates transformation of epithelial mesenchymal, then play a role in treatment of liver fibrosis.2. In vitro experimentIn vitro research Hep-G2 cells,use the TGF-β induced liver fibrosi s model in vitro, from immunofluorescence and Western Blot research to d iscuss the molecular mechanism of liver fibrosis of "Taohong qizhu ruan gan decoction".2.1 Immunofluorescence results show:Compared with the blank group, in model group E-cadherin expression decreased, Smad2, TGF-βR1, Vimentin expression enhanced;compare with the m odel group, treatment group can raise E-cadherin expression, reduce Smad2, TGF-βR1, Vimentin expression, that low dose group、middle dose group、hi gh dose group of "Taohong qizhu ruangan decoction", colchicine, Fuzheng Huayu recipe has good curative effect.2.2 Western Blot protein quantitative analysis shows that:Compared with the blank group, expression of E-cadherin of model grou p decreased, expression of Vimentin, TGF-βR1 rised, indicating that liv er fibrosis in vitro cell model building is successful, and enhancement of Smad2 expression, promote the TGF-βR1 expression, thus promoting the transformation of epithelial mesenchymal; Compared with model group, low, medium and high dose group of "Taohong qizhu ruangan decoction" can rais e E-cadherin expression, reduce Vimentin, TGF-βR1, Smad2 expression, medi um and high dose group of "Taohong qizhu ruangan decoction" have better curative effect,and colchicine,Fuzheng Huayu recipe has similar effect.In vivo experiments from blocking the histopathological changes of 1 iver fibrosis,liver tissue immunohistochemical protein expression, mRNA expression, and in vitro experiments Hep-G2 cells immunofluorescence and Western Blot, have reached a relatively consistent results:experience d ecoction "Taohong qizhu ruangan decoction" has markedly effect for trea tment of liver fibrosis. The results from one side interpret theoretical connotation of experience of prefessor Gui-zhi sun in treatment of live r fibrosis and prevention and treatment of liver cancer treatment. Characteristic and innovation of this subject:1. Introduce the modern bioinformatics technology, using the "scale-fre e network" and "topic model" technical to analysis 35 cases of primar y liver cancer with a history of liver fibrosis/liver cirrhosis, summed up the corresponding of "core syndrome" and "core" the formulas, the clinical study, the paper documents submitted by professor Gui-zhi sun corrects and interviews to investigate and review, for the treatment of liver fibrosis in prefessor Gui-zhi sun characteristics, etiology and pa thogenesis, syndrome differentiation, treatment strategies, sent the cli nical characteristic to carry on the deep analysis, puts forward the tre atment of liver fibrosis using gui-zhi sun teacher "Yiqi huoxue ruanjia n" "Buyi piwei" treating principles and core, the formulas for mining experience in old Chinese medicine to provide new way of thought.2. This study used the rat liver fibrosis model and Hep-G2 cell model, co mbining the experiments in vivo and in vitro, from liver fibrosis pathol ogical tissue morphology, E-cadherin, Vimentin, Smad2, TGF-β1,TGF-βR1 and RT-PCR and immunohistochemical protein gene mRNAexpression, traditionalCh inese medicine medicated serum in vitro were tested afterintervention in duced E-cadherin, Vimentin, Smad2 and TGF-βR1 immunofluorescence and West ern Blot protein expression, etc. discussing the famous elder doctor oftr aditional Chinese medicine treatment of liver fibrosis, prevention andtr eatment of liver cancer clinical experience is scientific connotation, h ace not seen similar reports.3. Professor Gui-zhi sun proposed that prevention of liver cancer is very important, based on the traditional Chinese medicine "prevention before disease", "cure disease early" and "prevention progress after disease", we should focus on control of diagnosis and treatment of liver fibrosis/ cirrhosis. Based on that and clinical medicine analysis, professorGui-zhi sun summed up the experience decoction "Taohong qizhu ruangan decoctio n", "Taohong qizhu ruangan decoction" contains Fried astragalus, atracty lodes, peach kernel, pink flower, chicken’s gizzard-membrane, and so on dru gs,original, belongs to the inheritance innovation.