OBJECTIVESTo study the quantitative relationship between the levels of serum liver fibrosis markers and fibrosis stages of liver tissues in patients with chronic hepatic diseases.

METHODSIn 118 patients with chronic hepatitis, fatty liver or cirrhosis, their Serum levels of LN, HA, PCIII and CIV were investigated by EIA and their liver histological changes were studied. The relationship between the levels of serum LN, HA, PCIII and CIV and the degrees of liver tissue fibrosis was analyzed quantitatively by using the SPSS11.0.

RESULTSA correlation between the levels of serum LN, HA, PCIII and CIV and the histologically assessed grades of inflammatory activity was found (r = 0.394, 0.449, 0.443, 0.351, respectively, P <0.01). The correlation between the levels of serum LN, HA, PCIII and CIV and the histological assessed stages of liver fibrosis was strong (r = 0.456, 0.564, 0.476, 0.421 respectively, P <0.01). The levels of serum LN, HA, PCIII and CIV of the patients with a stage 2 liver fibrosis were 110 ng/ml, 110 ng/ml, 100 ng/ml and 70 ng/ml respectively, with sensibilities of diagnosing stage 2 liver fibrosis at 70%, 79%, 79% and 74% respectively. Their specificities in diagnosing stage 2 liver fibrosis were 68%, 72%, 64% and 73% respectively. The levels of LN, HA, PCIII and CIV in serum of these patients diagnosing cut-off value in stage 4 liver fibrosis (early cirrhosis) were 130 ng/ml, 140 ng/ml, 120 ng/ml and 70 ng/ml respectively. Their sensibility of diagnosing liver cirrhosis was 79%, 93%, 79% and 86% respectively. Their specificity of diagnosing liver cirrhosis was 66%, 82%, 72% and 61% respectively. As shown by the ROC curves in these patients, differentiating patients with cirrhosis or without cirrhosis, serum HA level was more valuable than LN, PCIII, CIV (the areas under the curves = 0.938 vs 0.775, 0.787, 0.791 ) When serum HA was higher than 190 ng/ml, the veracity of diagnosing liver cirrhosis was 93%.

CONCLUSIONSThere is a certain quantitative relationship between the levels of LN, HA, PCIII and CIV in serum and the degrees of liver tissue fibrosis. The level of HA in serum is an important reference datum for early diagnosing liver cirrhosis.

Adolescent ; Adult ; Aged ; Child ; Fatty Liver ; blood ; complications ; Female ; Hepatitis, Chronic ; blood ; complications ; Humans ; Hyaluronic Acid ; blood ; Laminin ; blood ; Liver ; pathology ; Liver Cirrhosis ; blood ; etiology ; pathology ; Male ; Middle Aged ; Procollagen ; blood

No abstract available.
Humans ; Interferon-gamma* ; Pityriasis Rosea*

OBJECTIVETo investigate the clinical significance of the expression of serum differential protein in patients with chronic hepatitis B (CHB) related liver fibrosis.

METHODSOne hundred and ten CHB patients confirmed by liver biopsies were enrolled, 83 for modeling and 27 for verification. According to Ishak staging, 55 patients in the modeling group were with significant liver fibrosis ( F is more than or equal to 3 ) and 28 patients with normal/mild liver fibrosis ( F0-F2 ). While that in the verification group were 15 ( F is more than or equal to 3 ) and 12 ( F0-F2 ), respectively. MALDI-TOF-MS/MS was used to detect serum proteins and the spectrum for each sample was analyzed in FlexAnalysis3.0 to produce the spectrum of differential proteins. The results were compared with clinicopathologic diagnosis and the diagnosis model based on genetic algorithm was established and evaluated.

RESULTSThere were 15 proteins differentially expressed in significant liver fibrosis group and normal/mild fibrosis group ( P value is less than 0.01), in which the differences on proteins 2081.73 m/z and 1944.41 m/z were the most significant. Based on these two proteins, the coordinate system was set up and the diagnosis model based on genetic algorithm was established by six characteristic peaks. After detecting 12 cases of normal/mild liver fibrosis and 15 cases of significant liver fibrosis, the results showed that the diagnostic model could identify significant fibrosis ( F is more than or equal to 3 ) and normal/mild liver fibrosis ( F0-F2 ) at 100% recognition, 94.14% prediction and 100% accuracy.

CONCLUSIONSerum differential proteins examination can be used for early prediction of CHB related fibrosis. The study provides the basis for non-invasive diagnosis of hepatic fibrosis according to identifying the potential differences of the serum samples from patients with HBV related fibrosis.

Adult ; Blood Proteins ; analysis ; Female ; Hepatitis B, Chronic ; blood ; diagnosis ; pathology ; Humans ; Liver ; pathology ; Liver Cirrhosis ; blood ; diagnosis ; pathology ; Male ; Middle Aged ; Proteomics

OBJECTIVETo explore the protective mechanisms of glycine (Gly) on lipopolysaccharides (LPS) induced liver injury.

METHODSBABL/c mice were randomly divided into a LPS group, in which the animals were intraperitoneally injected with 10 mg/kg LPS, and a Gly group, in which the mice were pretreated with a 5% Gly-containing diet for 3 days before receiving the same dose of LPS. The livers of the mice were examined for histopathological changes. The TNF alpha and interleukin-10 (IL-10) levels in the blood plasma were measured using ELISA analysis. The mRNA expression of TNF alpha, IL-10 and Toll-like receptor 4 (TLR4) in hepatic tissues were detected using RT-PCR analysis. Protein expression of TLR4 in livers was detected using immunohistochemistry.

RESULTSThe Gly group mice had an improved survival rate and attenuated LPS-induced pathological changes in the liver tissues in comparison with those of the LPS group animals. The TNF alpha levels [(1,852.80+/-126.64) pg/ml vs (708.83+/-51.29) pg/ml, P<0.05] in plasma, as well as the expression of TNF alpha (A 1.59+/-0.14 vs. 0.91+/-0.11, P<0.05) and TLR4 (A 0.97+/-0.12 vs. 0.53+/-0.11, P<0.05) mRNA in liver tissues were decreased. However, the levels of plasma interleukin-10 [(344.09+/-31.70) pg/ml vs (418.64+/-38.86) pg/ml, P<0.05] were significantly increased and the peaking time left, shifted.

CONCLUSIONSGly pretreatment could attenuate LPS -induced liver injury in mice, which may be associated with its role in down-regulating TLR4 expression and up-regulating IL-10 production.

Animals ; Down-Regulation ; Female ; Glycine ; pharmacology ; Interleukin-10 ; blood ; metabolism ; Lipopolysaccharides ; adverse effects ; Liver ; drug effects ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; Toll-Like Receptor 4 ; metabolism ; Tumor Necrosis Factor-alpha ; blood ; metabolism ; Up-Regulation

BACKGROUNDSentinel lymph node (SLN) biopsy has become a common procedure for early breast cancer patients. The GeneSearch(TM) Breast Lymph Node (BLN) Assay is a real-time RT-PCR assay for the detecting nodal metastases larger than 0.2 mm. China Breast Cancer Clinical Study Group (CBCSG)-001a is a prospective multi-center clinical trial that was conducted to validate the GeneSearch(TM) BLN Assay in China.

METHODSThe SLNs from 90 consecutive patients were identified and dissected, and then sectioned along the short axis into multiple blocks. Intra-operatively, the odd blocks were tested by BLN assay and the even ones were used for frozen section, while all the blocks were evaluated by touch imprint cytology. Post-operatively, the remaining tissues were assessed by histological evaluation.

RESULTSA total of 189 SLNs was tested by BLN assay. The sensitivity, specificity, positive predictive value, and negative predictive value were 88.9%, 97.4%, 88.9% and 97.4%, respectively, for BLN assay, 75.0%, 100%, 100% and 94.4%, respectively, for frozen section, and 63.9%, 100%, 100% and 92.2%, respectively, for touch imprint cytology. The sensitivity of BLN assay was higher than that of touch imprint cytology (P = 0.01) and frozen section (P = 0.13). When assessing the nodes with micro-metastases, BLN assay had a significant higher sensitivity than frozen section (P = 0.023) and touch imprint cytology (P = 0.005).

CONCLUSIONThe GeneSearch(TM) BLN Assay is an accurate and rapid intra-operative assay for breast SLNs and it is suitable for application in general medical practice.

Adult ; Aged ; Breast Neoplasms ; complications ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; diagnosis ; Middle Aged ; Sentinel Lymph Node Biopsy ; methods

OBJECTIVETo explore the mechanism of endotoxin tolerance (ET) through observing the expression of interleukin 1 receptor associated kinase-4 (IRAK-4) during endotoxin tolerance development in Kupffer cells (KCs).

METHODSIsolated KCs of Balb/c mouse were divided into two groups: the non-endotoxin tolerance (NET) group and the endotoxin tolerance (ET) group, which were pretreated with 10 ng/ml lipopolysaccharide (LPS) for 24 h. Then, the two groups were treated with 100 ng/ml LPS. The expressions of IRAK-4 gene and protein level were determined by RT-PCR and Western blot. The activities of NF-kappaB of KCs and the TNFalpha level were estimated by ELISA at 0 h, 1 h, 3 h, 6 h and 12 h after LPS stimulation.

RESULTSThe ultimate level of IRAK-4, the activities of NF-kappaB and the TNFalpha level were evidently lower in the ET group than those in the NET group (t = 12.4, 17.4 and 138.9 respectively, P<0.01).

CONCLUSIONSPretreatment with LPS on KCs could induce endotoxin tolerance of KCs and inhibition of IRAK-4 expression may be one of the reasons for its development.

Animals ; Cells, Cultured ; Endotoxins ; immunology ; Immune Tolerance ; Interleukin-1 Receptor-Associated Kinases ; biosynthesis ; genetics ; Kupffer Cells ; cytology ; immunology ; metabolism ; Lipopolysaccharides ; immunology ; Male ; Mice ; Mice, Inbred BALB C

OBJECTIVETo investigate the relationship between the genotypes of hepatitis B virus and the clinical and liver pathological features of patients with chronic hepatitis in the Zhoushan Islands.

METHODSOne hundred eighty HBV DNA positive chronic hepatitis patients with HBV markers were enrolled in this study. They were at least second generation Zhoushan Island residents. One hundred forty-seven of them were males and 33 were females with an average age of 39.0+/-11.3. Among the 180 patients, 17 had ASC, 57 had mild CHB, 48 moderate CHB, 9 severe CHB, 6 SHB, 39 LC, and 4 had HCC. The genotypes of their serum HBV were detected by using PCR integrated with Tagman MGB probe technology, and their serum HBV markers, HBV DNA and liver functions were also examined. Out of 180 patients, 129 accepted a liver biopsy. A pathological evaluation was then performed.

RESULTSHBVs of genotype C, 135 cases (75.0%), of B, 40 cases (22.2%), and of B+C, 5 cases (2.8%) were found among these 180 patients. No genotype A or D HBV were found. The proportions of genotype C virus were 7/17, 86/114, 34/39, 6/6 in ASC, CHB, LC and SHB patients. In the hepatocellular carcinoma patients, there were 2 each of genotype B and C. Among the 99 patients with genotype C HBV, 84 cases (84.8%) showed moderate and severe inflammation histologically in their livers and among the 30 patients with B, 7 cases (23.3%) showed moderate to severe inflammation in their livers (z = 6.47, P less than 0.01). The proportion of genotype C HBV was significantly different from that of genotype B HBV in those that showed moderate and severe (S3-4) liver fibrosis. In patients infected with genotype C HBV who had moderate and severe liver pathological changes, their clinical manifestations reflected better the histological alterations of their livers.

CONCLUSIONGenotypes C, B and B+C HBV were found in CHB patients in the Zhoushan Islands of China, and type C was the predominant one. The liver pathological damage level of genotype C HBV infected patients is more serious than that of genotype B.

Adult ; China ; epidemiology ; DNA, Viral ; genetics ; Female ; Genome, Viral ; Genotype ; Hepatitis B virus ; classification ; genetics ; Hepatitis B, Chronic ; epidemiology ; pathology ; Humans ; Liver ; pathology ; Male ; Middle Aged

OBJECTIVETo explore the inhibitory effects on the activation of endotoxin-induced Kupffer cells (KCs) through short hairpin RNA (shRNA) targeting interleukin-1 receptor associated kinase-4 (IRAK-4) gene.

METHODSTwo effective transfection shRNA plasmid (pSIIRAK-4-A, pSIIRAK-4-B) and one invalidated plasmids (pSIIRAK-4-C) targeting IRAK-4 gene were constructed. The isolated mouse KCs were divided into three groups: the normal control group, the RNAi control group (pSIIRAK-4-C) and the RNAi effective group (pSIIRAK-4-A, pSIIRAK-4-B). Then KCs were stimulated with 0.1 microg/ml lipopolysaccharide (LPS) after 24 h transfection with the constructed plasmid. The expression of IRAK-4 gene and protein level were determined by RT-PCR and Western blot at 6 h after LPS stimulation, and the activities of NF-kappaB in KCs and the TNFalpha level were estimated by ELISA at 0 h, 1 h, 3 h, 6 h and 12 h.

RESULTSThe level of IRAK-4, the activities of NF-kappaB and the TNF-alpha level in the RNAi effective group were evidently lower than those in normal and RNAi control groups (P < 0.01) at 1 h, 3 h, and 6 h. Especially, the pSIIRAK-4-A group in which the changes of the above indices were of no difference (P > 0.05), had better inhibited effects than that of the pSIIRAK-4-B group (P < 0.01).

CONCLUSIONThe shRNA targeting IRAK-4 gene could effectively inhibit the activation of endotoxin-induced KCs.

Animals ; Endotoxins ; Interleukin-1 Receptor-Associated Kinases ; genetics ; metabolism ; Kupffer Cells ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; RNA Interference ; RNA, Small Interfering ; genetics ; Signal Transduction ; physiology

OBJECTIVETo explore the possible mechanism and optimal treatment phase of glycine for inhibition lipopolysaccharide (LPS) induced Kupffer cells (KCs) activation.

METHODSThe KCs were isolated from 40 BALB/c mice and divided into four groups: the endotoxin group, the prevention group, the early treatment group and the later treatment group (n = 10). The endotoxin group was treated with 10 mg/L LPS, and in the other three groups, glycine (1 mmol/L) was given 24 h before, or at 0 h or 4 h respectively after LPS stimulation. At 0 h, 1 h, 2 h, 6 h and 12 h after LPS stimulation, the mRNA levels and protein expression of interleukin-1 receptor associated kinase-4 (IRAK-4) were determined by reverse transcription polymerase chain reaction (RT-PCR) and Western blot respectively, and nuclear factor-kappaB (NF-kappaB) activities as well as tumor necrosis factor alpha (TNF-alpha) levels were also detected by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe climax values of IRAK-4, NF-kappaB and TNF-alpha were significantly higher in the endotoxin group and the later treatment group than that in the other two groups (t = 3.17, 4.33, 2.47, 126.73, P < 0.01).

CONCLUSIONThe results indicated that prophylactic or simultaneous treatment with glycine could effectively inhibit LPS-induced KCs activation by inhibiting IRAK-4 expression.

Animals ; Cells, Cultured ; Drug Interactions ; Glycine ; administration & dosage ; pharmacology ; Interleukin-1 Receptor-Associated Kinases ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Kupffer Cells ; drug effects ; metabolism ; Lipopolysaccharides ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; NF-kappa B ; metabolism ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Time Factors ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo detect the presence of endothelial injury in patients with severe acute respiratory syndrome (SARS) via enhanced levels of tissue-type plasminogen activator (t-PA) and soluble thrombomodulin (sTM).

METHODSCase patients were from Xuanwu Hospital (Capital University of Medical Sciences, Beijing, China), and all of them met clinical criteria for SARS. Healthy controls were some of the hospital employees. Endothelial injury bio-markers tPA and sTM were detected by commercial ELISA-methods.

RESULTSClassic plasma markers of endothelial injury, tPA and sTM significantly elevated in SARS patients in comparison to controls [t-PA: 1.48 +/- 0.16 nmol/L versus 0.25 +/- 0.03 nmol/L (P<0.0001), and sTM: 0.26 +/- 0.06 nmol/L versus 0.14 +/- 0.02 nmol/L (P<0.05)]. The only patient who died had extremely high levels of these endothelial injury markers (t-PA: 2.77 nmol/L and sTM: 1.01 nmol/L). The likelihood ratio analysis indicated the excellent discriminating power for SARS at the optimal cut-point of 0.49 nmol/L for tPA and 0.20 nmol/L for sTM, respectively. Significant numerical correlations were found among these endothelial injury markers in SARS patients. The numerical coefficient of correlation Pearson r between t-PA and sTM was 0.5867 (P<0.05).

CONCLUSIONIncreased plasma concentrations of tPA and sTM in patients with SARS suggest the possibility of endothelial injury. SARS patients might need anticoagulant therapy or fibrinolytic therapy in order to reverse intraalveolar coagulation, microthrombi formation, alveolar and interstitial fibrin deposition. It may not only provide a useful treatment and prognostic index but also allow a further understanding of the pathological condition of the disease.

Adult ; Biomarkers ; blood ; Case-Control Studies ; China ; Female ; Humans ; Male ; Prognosis ; Severe Acute Respiratory Syndrome ; blood ; Thrombomodulin ; blood ; Tissue Plasminogen Activator ; blood