| ObjectivePrimary liver cancer (PHC) is a kind of disease with high incidence and mortality. It is a serious threat to human health. The majority of patients with liver cancer are associated with the infection of hepatitis B virus (HBV). The liver cirrhosis with HBV infection, especially intercurrent with dysplastic nodule, have high probability of developing into liver cancer. It has been regarded as the hepatic precancerous lesion. In recent decades, although PHC treatments have been developed and perfected, but its clinical efficacy has not been significantly improved. Therefore, it has a very important practical significance to prevent or delay the formation and development of liver cancer by interfering in the stagy of precancerous lession. Based on tutor's clinical and experimental work, we tried to do a preliminary study about the syndrome law of TCM in the ascites with hepatitis B cirrhosis (intrahepatic nodular>1cm), and to investigate the mechanism of prevention and treatment of hepatic precancerous lesion with Kangxianyiai Decoction by Means of observing the relationship between hepatocyte growth factor(HGF) and its receptor(c-Met) and alpha smooth muscle actin(α-SMA).Methods1 Clinical research Based on extensively reviewing the relevant literature, collected the clinical symptoms of TCM of the ascites with hepatitis B cirrhosis, then established an initially clinical epidemiological survey questionnaire. At last, determined the required clinical symptoms of the survey questionnaire item pool in TCM, and made the corresponding classification of quantitative criteria symptoms by relevant expert advice and feasibility analysis, repeated discussions and seminars and clinical research guidelines. Used uniform questionnaire to collect 134 cases clinical syndromes data of the ascites with hepatitis B cirrhosis (intrahepatic nodular>1cm). We use the method of factor analysis to investigate its characteristics of TCM syndrome, and to discuss the relationship between the objective indexes with the characteristics of TCM.2 Experimental research Established the animal models of diethylnitrosamine(DEN)-induced hepatic precancerous lesion, and used Kangxianyiai (KXYA) Decoction to treat. Fufangbiejiaruangan(BJRG) tablets and colchicine tablets were taken as control drug. Compared the difference of rats in each group on the general situation and the liver pathological change, and determined the expression levels of glutathione-S-transferases-π(GST-Pi), of proliferating cell nuclear antigen(PCNA), alpha-fetoprotein(AFP), AFP-L3,α-SMA, HGF and c-Met in the liver of the rats in each group by using immunohistochemistry, Enzyme-linked immunoassay(ELISA) and real-time fluorescence quantitative PCR (RQ-PCR), so as to reveal the mechanism of intervention on hepatic precancerous lesion by Kangxianyiai decoction.Results1The results of clinical research Used collected clinical data of 134 cases of the ascites with hepatitis B cirrhosis (intrahepatic nodular>lcm) to do the frequency analysis derived:The peak age incidence of the ascites with hepatitis B cirrhosis (intrahepatic nodular>lcm) was between 51 and 60 years old; the time suffering from liver cirrhosis less than five years occupied 67.16%; the proportion that the history of ascites appears less than five years was 85.07%; 62.69% patients with ALB value lower than normal, and the ascitic depth of right lateral position less than 3cm was the highest frequency, accounted for 35.82%. By means of the unsupervised factor analysis to analyze the collected clinical symptoms, the characteristics of TCM syndrome of the ascites with hepatitis B cirrhosis (intrahepatic nodular>1cm) can be summarized as:the syndrome of dampness and heat in the liver and the gallbladder with spleen deficiency, the syndrome of stagnation of liver qi and deficiency of the spleen with water retention, the syndrome of fire due to qi stagnation with blood stasis, the syndrome of qi deficiency and blood stasis, the syndrome of qi and damp stagnation, the syndrome of stagnation of liver-qi, the syndrome of damp pathogen obstructing the spleen, the syndromes of dysfunction in transport due to spleen deficiency, the syndromes of insufficiency of the liver and the kidney with internal block due to blood stasis. The correlation analysis between syndrome characteristics and clinical data obtained has being done. The factors such as antiviral therapy affect the distribution of syndrome characteristics.2The results of experimental research On the general condition, the model group begun to present some pathological behaviors after pouring the DEN into stomach for three weeks. Every treatment group had such similar condition but was lighter than the model group. In the twelfth week, the rats in every group died in succession except the negative control group. At the end of the experiment,6 rats in the model group were dead while 2 rats in KXYA group died (one of the rats was due to the careless drug delivery in the second week).4 rats in the BJRG group were dead and 4 rats in the group of colchicin died too. In conclusion, the fatality rate was lowest in the KXYA group. On the Gross morphology, liver surface was rough, and diffuse nodules formed in model group. It was found by pathological examination that liver cell necrosis, collagen fibers hyperplasia obviously, false lobular formed widely, liver cell volume increased, and the nucleus was pleomorphic. The pathological performances of liver of the BJRG group and the colchicine group were similar to the model group's. There were no obviously rugosity in the liver surface of the KXYA group, and the nodules were smalerl than these of model group. Masson's trichrome stain showed that the condition of liver fibrosis in each rat group was similar with HE's results, but with more obvious the coloring of collagen fibers. The results of immunohistochemistry showed that:Compared with the model group, the KXYA group significantly reduced the GST-Pi positive foci area and number, while also reducing the PCNA-positive cell labeling index, and the expression levels of a-SMA, which has a remarkable statistical significance(P<0.01). The results of ELISA showed that: Compared with the model group, the KXYA group can significantly reduce the expression levels of AFP and AFP-L3, which has a remarkable statistical significance(P<0.05 and P<0.01); the ratio of AFP-L3 and AFP is also lower than the model control group, but the difference has no statistical significance(P>0.05). The results of RQ-PCR showed that:There were significant difference in the relative expression levels of HGF mRNA between KXYA group and model control group(P<0.05). Kangxianyiai Decoction can significantly increase the expression level of HGF mRNA in rat hepatic precancerous lesion. c-Met mRNA were all low expression in the each group of DEN-induce hepatic precancerous lesion, and the difference in each group has no statistical significance(P>0.05).Conclusionsthe characteristics of TCM syndrome of the ascites with hepatitis B cirrhosis (intrahepatic nodular>1cm) were:the syndrome of dampness and heat in the liver and the gallbladder with spleen deficiency, the syndrome of stagnation of liver qi and deficiency of the spleen with water retention, the syndrome of fire due to qi stagnation with blood stasis, the syndrome of qi deficiency and blood stasis, the syndrome of qi and damp stagnation, the syndrome of stagnation of liver-qi, the syndrome of damp pathogen obstructing the spleen, the syndromes of dysfunction in transport due to spleen deficiency, the syndromes of insufficiency of the liver and the kidney with internal block due to blood stasis. The characteristics of TCM syndrome have relativity with antiviral therapy, PT, WBC.It's effective to reduce the extent of liver cell necrosis, collagen fibers hyperplasia and liver cell dyplasia, delay the occurrence and development of hepatic precancerous lesion by Kangxianyiai Decoction. The mechanism of action related to increased expression of HGF mRNA, and inhibited activation of hepatic stellate cells (HSC), promoted degradation of extracellular matrix(ECM).
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