| BackgroundHepatitis C is caused by hepatitis C virus(HCV)infection.It is a small RNA virus belonging to the Flaviviridae family.American scientists cloned the hepatitis C virus gene in 1989.HCV is mainly transmitted through blood transfusion,acupuncture,drug addiction and mother-to-child transmission.According to statistics from the World Health Organization,approximately 180 million HCV infected person all over the world with an infection rate of about 3%,and about 35,000 new cases each year.At present,HCV has shown a global epidemic trend,which can not only cause chronic hepatitis and liver fibrosis,and eventually progress to cirrhosis.Among them,about 3-10% can also develop to hepatocellular carcinoma(HCC).This shows that hepatitis C has become one of the important diseases that seriously threatens human health and is worthy of in-depth research.At present,the clinical diagnosis of hepatitis C is mainly based on viral serological tests,supplemented by pathological diagnosis.The light microscope can only be diagnosed by the inflammatory response and the degree of lymphatic infiltration in the leaflet and the manifold area,which is not obvious at early stage.The observation and summary of the ultrastructure under the electron microscope has become the main breakthrough point of pathological diagnosis.Not only can early pathological diagnosis be made through changes in the ultrastructure of cells,but also the mechanism and prognosis of disease development can be speculated.However,there are fewer studies on the ultrastructure of hepatitis C.The present study focused on the conventional pathology and ultrastructure of hepatitis C,observed the characteristics of each stage at the ultrastructural level,provided a basis for further study of ultrastructure of hepatitis C,and provided a pathological basis for its clinical staging and treatment,and provided the study basis of microstructure.Research purposesThis study performed hematoxylin and eosin(HE)staining and special staining on the liver tissues of patients with hepatitis C with different degrees of inflammation,and evaluated the pathological stages according to the pathological changes.Meanwhile,the present study observed the ultrastructure of hepatocytes in patients with hepatitis C of different stage and cancerization,and explored the relationship between the hepatocyte ultrastructural characteristics of hepatitis C and cancerization,and then predicted the development and prognosis of hepatitis C patients.Research methods1.Research subjects: a total of 120 patients in the 900 th Hospital of the Joint Logistics Support Unit which the virus serological test for HCV antibody is positive collected from August 2011 to August 2017,of which 4 cases developed into HCC after 3 years follow-up.Percutaneous liver biopsy was performed to collect the liver tissue specimens of these patients.20 patients were randomly collected the liver tissue around hepatic hemangioma as a negative control.2.Data collection and processing: the basic information and clinicopathological data of the above cases was collected,including the patient’s gender,age,viral serological detection of viral infection,whether it is the first attack,liver fibrosis assessment,postoperative treatment plan and prognosis follow-up work.After HE staining,collagen fiber staining and mesh fiber staining,the tissue morphology was observed under an optical microscope,and the classification was carried out according to the modified Scheuer histological scoring system.3.Using transmission electron microscope(TEM)on the basis of light microscope diagnosis,using liver biopsy tissue extracted from paraffin-embedded tissue,and using gasoline deparaffinization instead of xylene deparaffinization for TEM sample preparation,the ultrastructural characteristics of hepatitis C with different degrees of inflammation and cancerization was observed and summarized.Result1.Observation results under light microscope: 120 cases of chronic hepatitis C includes 56 cases of mild chronic hepatitis C,45 cases of moderate chronic hepatitis C and 19 cases of severe chronic hepatitis C hepatitis.In mild microscopic hepatitis C,hepatocyte edema and degeneration,hepatocyte ribosylation,and occasional spot necrosis was observed;slight inflammation of the confluent area,follicular formation was observed,without fibrosis or little fibrous tissue hyperplasia in the confluent area.In moderate chronic hepatitis C,hepatocyte edema and degeneration,punctate necrosis,and interfacial inflammation was observed;and inflammation in the manifold area,fibrous tissue hyperplasia,and may be accompanied by the formation of fibrous septa.In severe chronic hepatitis C,hepatic necrosis and significant interface inflammation with inflammation in the confluent area accompanied by the formation of lymphatic follicles was obvious;and significant fibrous tissue hyperplasia,bridging or arcuate fiber and early pseudolobular formation.2.Tissues were extracted from paraffin block and deparaffinized by gasoline.Under TEM,the hardness of the embedded block was moderate,the effect of soaking was ideal,the process of section preparation was smooth,the staining contrast was obvious,the ultrastructure of liver tissue was clearly identifiable,the internal structure of the cell and organelles were intact,and the observation results were satisfactory.3.Observation results under TEM: among 56 cases of mild chronic hepatitis C,14 cases showed hepatocyte cytoplasmic glycogen increase and aggregation distribution;20 cases showed normal cytoplasmic rough surface in the cytoplasm decreased cytoplasmic reticulum and increased endoplasmic reticulum on the slip surface.Among 45 patients with moderate chronic hepatitis C,10 cases showed increased intracytoplasmic lipid droplets with large lipid droplet formation;38 cases showed increased intercellular collagen fibers.Among 19 patients with severe chronic hepatitis C,12 cases showed mitochondrial deformation,significantly increased lipid droplets and intercellular collagen fibers,and 3 cases showed partial mitochondrial hypertrophy,mitochondrial crest rupture,and vacuolation of endoplasmic reticulum.4 patients with cancerization: the endoplasmic reticulum was obviously cystic dilated,the mitochondria decreased,and the mitochondrial vacuoles changed.The results showed that there was a statistical correlation between the increase of glycogen,decreased rough endoplasmic reticulum,mitochondria fusion,increased lipid droplets,the intercellular collagen fibers,endoplasmic reticulum vacuole and the degree of hepatitis C lesions(P <0.05).Conclusion1.We improved the traditional xylene deparaffinization method by using gasoline deparaffinization.Observation results under TEM showed that gasoline deparaffinization was more complete,ultrathin slices were more intact,paraffin-embedded tissue were used more effectively by gasoline deparaffinization,the ultrastructure of organelles in hepatocytes was preserved intact,the form was clearly recognizable,and the effect of dyeing was ideal,the range of electron microscope specimens was expanded,a bridge of communication between light microscope and electron microscope diagnosis of the same specimen was established,and the trouble of secondary clinical sampling due to the need of electron microscope diagnosis was avoided.Gasoline deparaffinization saved the time of sample preparation,greatly improved efficiency of microscopy production,guaranteed the observe result,improved review speed and diagnostic accuracy,was less damage to the operator and environmentally friendly,and was worth of widely use.2.Observed under TEM,the hepatocytes ultrastructure of chronic hepatitis C will change accordingly with the degree of inflammation and fibrosis.Hepatocytes of hepatitis C with different degrees of inflammation have characteristic ultrastructural changes,which can be used to predict the development and prognosis of hepatitis C.3.Furthermore,it has certain significance to assist in the clinical staging and to predict the development trend of the hepatitis C virus through the observation of characteristic changes in ultrastructure. |
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