| 1Background and AimsHepatitis B virus (HBV) infection, a major health problem worldwide, is thought to be one of the main causes of liver-related diseases. A part of patients with chronic hepatitis B (CHB) may progress to liver failure due to severe acute exacerbation. At present, no sensitive and reliable criteria or system is available for clinical evaluation of liver failure. Disease progression in chronic hepatitis is often more hidden and difficult to be recognized by current diagnostic methods and techniques, easily overlooked by both doctors and patients. The diagnosis and treatment until clinical symptoms during the course often has entered end stage, which increases the difficulty of treatment. Furthermore, the prognosis is poor. What’s the worse, the costs of treatment is increased, adding to the economic burden of patients’ family and society, bringing the unstability of the society. At present, China is still lack of senstitive and reliable indicators of clinical evaluation or system for liver failure. So, finding a sensitive and reliable indicator of clinic which is easy to detect will be very valuable fo determining the severity of the disease.Macrophages play an important role in normal homeostasis and during various pathological conditions such as infection,(chronic) inflammation,atherosclerosis and cancer. CD163(Cluster of Differentiation163) is a highly expressed macrophage membrane protein belonging to the scavenger receptor cysteine rich (SRCR)domain family. Soluble CD163(soluble CD163, sCD163) is readily detectable in the serum,and tissue fluid and t can also reflects the activation of monocyte-macrophage.The most clearly described function of CD163is to mediate macrophage to clear redundant free hemoglobin in human body as the receptor of Hb-Hp complex. The potential functions of sCD163are quite extensive. If these functions are further excavation and concretely applied to the study of liver failure, itwould be very meaningful. Nowadays, China’s current clinical and pathological studies of liver failure is not perfect. Is sCD163associated with liver failure, and what is the relationship between sCD163and inflammation? These questions are both worth exploring.In scientific experiments and clinical testing often encountered specimens failed to timely submitted. And the temperature for delivery cannot be low, or the delicery requires multiple test cases. Thus, the affecting factors of the storage of specimens for the stability of our experimental results are well worth exploringMy objective is to investigate the diagnostic values of sCD163in patients with severe hepatitis B or various inflammations.2Materials and Methods2.1Materials2.1.1Serum collectionThe serum of100patients who stay in the First Affiliated Hospital Zhejiang University were collected. Then they were divided into2groups, with50cases in each group. They can be called group A and B. Group A were also divided into2groups and kept at25℃and4℃respectively. The two groups were kept-80℃for3,9,24,48,72hours and7days. Group B can be called repeated freezing-thawing group,, which was frozen and thawed for0,1,2and3times.Serum samples used in this study were collected, including38liver failure patients,40hepatitis B virus-induced liver cancer patients (HBsAg positive, HCC),40hepatitis B virus-induced hepatic cirrhosis patients (HBsAg positive,LC),38chronic hepatitis B patients(CH),40hepatitis B virus carrie(ASC),40fatty liver patients without hepatitis B virus infection(SS)38chronic glomerulonephritis patients (CGN),38community acquired pneumonia patients (CAP),38acute pancreatitis patients (AP) and40healthy subjects served as control.2.2MethodsThe concentrations of sCD163was detected by ELISA.2.4Statistical analysisContinuous data are expressed as the mean±standard deviation and were analyzed using Student’s t-test. Categorical data were expressed as counts and percentages and analyzed by the chi-square test. Correlations among the study variables were tested by Pearson’s correlation coefficient. P<0.05was demonstrated significant different.3Results3.1The influence of different storage conditions on sCD163ELISA test results3.1.1There were no significant differences on the influence of sCD163level for samples which were reserved at25℃for3,9,24,72hours and7days (P>0.05).3.1.2There were no significant differences on the influence of CD163level for samples which were reserved at4℃for3,9,24,72hours and7days (P>0.05).3.1.3There were no significant differences on the influence of sCD163level for samples which were reserved at25℃and4℃(P>0.05). 3.1.4The impact of repeated freezing-thawing which was exerted on sCD163ELISA test resultsThe number times of freeze and thaw also has no effect on the level of CD163(P>0.05).3.2Expression of sCD163in patients with liver disordersThe expression of sCD163was remarkably elevated in patients with liver failure compared with that of the control, and patients with liver failure, hepatitis B virus-induced liver cancer, hepatitis B virus-induced hepatic cirrhosis, chronic hepatitis B, hepatitis B virus carrier, and fatty liver, respectively(P<0.05). The expression of sCD163was significantly decreased in the control group compared with those in patients withliver failure、hepatitis B virus-induced liver cancer、 hepatitis B virus-induced hepatic cirrhosis、 chronic hepatitis B、hepatitis B virus carrier, and fatty liver, respectively (P<0.05).3.3Expression of sCD163in various inflammatory diseasesCompared to the expression of sCD163in the control group, sCD163levels in patients with chronic hepatitis B, chronic glomerulonephritis, community acquired pneumonia, and acute pancreatitis were significantly increased (P<0.05). However, no statistical differences were observed between the sCD163expression in the patients with chronic hepatitis B, chronic glomerulonephritis, community acquired pneumonia, and acute pancreatitis (P>0.05).3.4The correlation between sCD163and clinical parameters of liver failure patients, including Alanine transarninase (ALT), glutamic-oxalacetic transaminase (AST), prothrombin time (PT), international normalized ratio (INR), total bilirubin (TBIL) and e-antigen (e-AG). The results revealed that no correlation was noticed between sCD163level and these parameters. This indicated that those clinical parameter proteins showed no effects on the secretion of sCD163.4Conclusions4.1The repeated freezing-thawing has little influence on the result of ELISA for sCD163level in serum of patients. It has little influence on the result of ELISA for sCD163level when the samples were reserved at4℃and25℃within7days.4.2sCD163could be used as a biomarker for the diagnosis of liver failure. Also, it could be used for the evaluation of prognosis of liver failure, and the up-regulation of sCD163was closely correlated with the liver failure. In addition, sCD163showed no organ-specificity in patients with hepatitis, chronic glomerulonephritis, pneumonia, and pancreatitis. |
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