Study Of Immunohistochemical Expression Of C3d In Glisson's Capsule And Fibrous Septa Within Normal And Hepatitis B Cirrhosis Of Human Liver Tissues

[Objectives]1. To observe expression of C3d in Glisson's capsule (GC) within normal human liver tissue, and and to explore the reason, mechanism and significance of which of the deposition.2. To observe the expression of C3d in fibrous septum (FS) and GC within 76 hepatitis B cirrhosis (HBC) cases and 74 cases hepatitis B cirrhosis associated with hepatic carcinoma (HBC-HC), respectively, and to explore the source of FS fibers.3. To observe the inflammatory pathological characteristics of HBC and HBC-HC specimens, and to explore whether there are any relationship with follow-up and functional parameters.4. Observe the pathological features of portal activity (PA), and to explore its cinicathological significance. PA means the inflammation of or within GC wall.[Methods]1. PartⅠ60 archived paraffin-formed normal liver specimens were obtained from the Department of Pathology, Fuzhou General Hospital of Nanjing Military Command of Chinese PLA from 2006 to 2010. All specimens were performed HE, reticular fiber-Masson staining and immunohistochemistry for IgG, IgM, IgA, C3d, C1q, and Fib staining, only C3d and CD21 staining for specimens of other organs of control group. Two cases of fresh normal adult liver tissues were choosed randomly to explore the C3d immune electron microscopy staining.2. PartⅡ150 cases of archive paraffin specimens of liver transplant recipients, including 76 cases of Hepatitis B Cirrhosis (HBC) and 74 cases of Hepatitis B Cirrhosis associated with hepatic carcinoma (HBC-HC), were obtained from the Department of Pathology, Fuzhou General Hospital of Nanjing Military Command of Chinese PLA from 2008 to 2010. The subtype of liver cancer was 70 cases of HCC vs4 cases of intrahepatic cholangiocarcinoma. All specimens were performed HE, reticular fiber-Masson staining and immunohistochemistry with HBSAg, HBCAg, IgG, IgM, IgA, C3d, C1q, Fib and CD21 staining.10 cases of each group were choosed randomly to explore characteristics of the reticular fiber-C3d and electron microscopy staining.[Results]1. PartⅠ1.1 It displayed that deposition of C3d within fiber layer of liver capsule, GC hepatic vein adventitia in varying degrees in normal liver tissue, mainly in the much more dense bundles of collagen fibers. 83% (50/60) cases of GC were strong positive for C3d, and C3d-positive collagen fibers were around the interlobular artery and vein and the periphery of the portal.1.2 The comparison among the normal liver group: 21~70-year-old group, 3~20-year-old group, and <3-year-old group, there were significant differences of the expression of C3d in liver capsule, GC, arterial intima, adventitia of hepatic vein, sinusoidal endothelial. The expression of C3d in 21~70-year-old group and to～20 age group was significantly higher than the <3-year-old group (P<0.001), also the expression of C3d in GC and the hepatic venous adventitia in 21~70-years-old group was significantly higher than the 3~20-year-old group (P<0.05).1.3 The expression of C3d in hepatic blood sinus: lobular sinusoidal endothelial cells in some specimens revealed a small amount of expression of IgG (26/60 cases, PA = 0.82±0.87) or C3d (17/60 cases, PA = 0.32±0.55), which mainly attached to the 1 or 2 area of lobules.1.4 CIEM staining: colloidal gold particles unevenly scattered deposited between the lacertus of collagen fibers in GC, but not closely integrated with the collagen fibers, the hepatic artery and liver sinusoidal endothelial cellular membrane also demonstrated a small amount of deposition of C3d.1.5 The expression of C3d in spleen: 75% of spleen capsule, spleen trabecular fibrous sheath showed the C3d's deposition, which deposited mainly in the much denser collagen lacertus. Trabecular central artery and splenic artery intima also disclosed the strong expression of C3d (30/40 cases, 75%). A variety light zone of germinal center of lymphoid follicles in lymphoid tissue showed the strong positive of C3d, with irregular network (61/73 patients, 83.56%).1.6 The comparison among the normal splenic groups: 21~70-years-old group, 3~20-year-old group, and <3-year-old group, there were significant differences of the expression of C3d in the splenic capsule, fibrous sheath of spleen trabecular trabecular artery intima, the central artery intima, lymphoid follicles. The expression of C3d in 21~70-years-old group, 3~20-year-old group was significantly higher than <3-year-old group (P<0.001).2. PartⅡ2.1 The group 60.67% (91/150) false lobular fibrous septa were C3d positive and 63.33% (95/150) GC was C3d-necgtive. C3d deposited mainly in the dense fibers of collagen fiber of FS and GC. C3d-positive area of collagen fibers had no edema, inflammatory cell infiltration. C3d-positive collagen fibers of FS and GC were continued with each other. C3d-negative FS of pseudolobules and GC showed the edema, inflammatory cell infiltration, as the texture of fibrous tissue was loose, also FS and GC was no continuity in the structure.2.2 In each subgroup of HBC and HBC-HC groups: the average of C3d in C3d-positive cases in GC of pseudolobules and GC of the large nodular cirrhosis were higher than that the mall nodular liver cirrhosis, the difference was significant (P<0.005).. The average of C3d in C3d-positive cases in FS of pseudolobules and GC of the mixed nodular cirrhosis was between large and small nodule cirrhosis groups, the difference was significant (P<0.005).150 cases of cirrhosis of the liver tissue, there were significant differences in pathological features between C3d-positive group and negative group. Chronic hepatitis stage G and portal activity (PA) score in the C3d-negative group was significantly higher than that of C3d-positive group (P<0.001), and T lymphocytes, B lymphocytes and plasma cells were higher than C3d negative group. Lymphocytic aggregation and lymphoid follicles were observed in some cases, and the rate of lymphoid follicles in C3d negative group was significantly higher than C3d-positive group (P<0.001).2.3 For the HBC and HBC-HC group, the false lobular fibrous septa and GC with C3d-positive group indicated that the liver function tests were significantly lower than C3d negative group, which the difference was significant (P<0.001).2.4 Reticular fiber-C3d staining showed the fibers of FS sprout from the GC, which was C3d positive liver cirrhosis, with the positive mode similar to the GC.2.5 TEM observation showed that there was no deposition of immune complexes in FS and GC in liver cirrhosis tissues.[Conclusions]1. The deposition of C3d in GC in normal liver tissue, normal spleen trabecular fibrous sheath and small arterial intima beyond 3-year-old, which represent a normal physiological phenomenon.2. C3d can be used as an immune marker of GC fiber of normal liver tissue.3. The portal area fibrosis and centrilobular fibrosis are two types of different mechanisms of liver cirrhosis, we believe that the essence of portal area fibrosis is GC fibrosis; otherwise C3d can be used as a immune marker of GC-oriented fibrosis.4. FS and GC stained with C3d in the cases that manifested C3d negative often accompanied by obvious active inflammation, higher aminotransferase and total bilirubin, and faster progression of disease.5. PA is a specific pathologic manifestnation of some hepatitis, which emergence often prompt the active hepatitis.