Biliary atresia (BA) is one of the most serious digestive system diseases, which threatens the health of infants. Liver fibrosis is a major cause of death in children with BA. In the process of the pathogenesis of BA, virus infection can in?duce a series of immune and inflammatory reaction, result in a decrease of regulatory T cells (Treg cells) and high expression of CD14, activating a variety of inflammatory pathways and TGF-β/Smad2/3 pro-fibrogenic pathway, which produces a large number of medium damage of liver cells and bile duct cells, releases proinflammatory factor, oxygen metabolism matter and cytokines. These changes further aggravate damage of hepatobiliary system and cause the internal environment imbalance of liver parenchyma cells. The imbalance of internal environment with adaptive degeneration and necrosis in liver parenchyma cells, hepatic macrophages and gathered inflammatory cells leads to the activation of hepatic stellate cells (HSCs). HSCs can be converted into fibroblast cells, and promote the process of liver fibrosis. Immune and inflammatory lesions, pro-fibrogenic pathway are the important factors in contributing to liver fibrosis and cirrhosis of biliary atresia.

Objective:The clinicopathological features, diagnosis, and prognosis of follicular thyroid carcinoma (FTC) with distant me-tastasis as the first manifestation were evaluated in this study. Methods:A total of 129 FTC cases with clinical data were retrospective-ly analyzed in the Department of Pathology, Tianjin Medical University Cancer Institute and Hospital (January 2001 to January 2016). Survival analysis and conjoint analysis on FTC with clinical data, diagnosis, and morphological characteristics with distant metastasis as the first manifestation were performed. Results:Among the 129 FTC cases, 24 cases demonstrated distant metastasis as the first mani-festation (18.6%). Bone metastasis was the most common (13.2%). The presence of mass and pain at the metastatic sites were the usu-al clinical complaints. The morphological characteristics of FTC with distant metastasis can be classified into four subtypes:microfollicu-lar (10 cases), solid (4 cases), normofollicular (9 cases), and macrofollicular (1 case). Immunostaining tests on thyroglobulin and thyroid transcription factor-1 showed positive results in FTC with metastasis. Survival analysis showed that the five-year survival rates in the 24 cases were 87.1%. The prognosis of patients with solitary metastasis was better than that of patients with multiple metastasis (P=0.022). A higher survival rate was found in the normofollicular and macrofollicular subtypes than that detected in the microfollicular and solid subtypes (P=0.012). Conclusion:FTC is susceptible to distant metastasis. Some patients with FTC demonstrated distant me-tastasis as the first manifestation, and their diagnosis can be confirmed by pathological feature analysis and immunostaining. The prog-nostic significance is possibly related to the number of lesions of FTC with distant metastasis and histopathological subtypes.

Objective To investigate the expression and function of transforming growth factor (TGF)-β1 and Smad2 in liver fibrosis of biliary atresia (BA). Methods Liver biopsy specimens were collected from autopsy (normal group, n=5), congenital biliary dilatation (CBD group, n=10), BA patients underwent Kasai procedure (early hepatic fibrosis group, n=19) and liver transplantation (transplantation group, n=11). The first three groups were collected from January 2010 to July 2014 in Tianjin Children’s Hospital, and the last group was collected from January 2013 to January 2014 in Tianjin First Central Hospital. The hematoxylin and eosin (HE) stain were used to observe the degree of liver fibrosis of four groups. Immunohistochemistry (IHC) was used to observe expressions of TGF-β1 and Smad2 in liver tissues of these samples. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to test the quantitative mRNA of TGF-β1 and Smad2 in these samples. Results Results of HE showed that no fibrosis in autopsy group, mild fiber cell hyperplasia in CBD group, severe fibrosis in Kasai group and significant pseudolobule in transplantation group. Results of IHC showed that TGF-β1 was expressed in the cytoplasm of hepatocytes, bile duct cells, lymphocytes and neutrophils. The average optical density of TGF-β1 was the highest in Kasai group compared with that of other three groups (P < 0.05). There was no significant difference in Smad2 expression in cytoplasm of hepatocytes, bile duct cells and lymphocytes between four groups (P>0.05). Results of qRT-PCR showed that both TGF-β1 mRNA and Smad2 mRNA were the highest in early hepatic fibrosis group than those of CBD group and transplantation group (P<0.017). Conclusion In early stage of BA, TGF-β1 and Smad2 promote liver fibrosis until the formation of P-P,P-C desmosome structure. However, with BA fibrosis becomes more serious, the pro-fibrogenic function of TGF-β1 and Smad2 becomes less.

Objective To analyze the impact and its mechanism of down-regulation of AnnexinA2 expression on prostate cancer(PCa) invasion and metastasis.Methods The expression of AnnexinA2 in three prostate cancer cell lines with different metastasis ability,including LNCaP (lower metastasis ability),PC3 (lower metastasis ability),C4-2B (higher metastasis ability) were detected by Western blot.The correlations between the expression of AnnexinA2 and the metastasis ability of prostate cancer cells were also evaluated.The siRNA was used in PC3 cells to down-regulate the expression of AnnexinA2,the cell proliferation assay was performed by MTT method,the cell apoptosis level was detected by flow cytometry,the expression of MMP-2 and MMP-9 were detected by Westrn blot,the in vitro invasiveness of PC3 was detected by transwell cabinet test,and the migration ability of PC3 cells was detected by scratch test,respectively.Results The grayscale value of AnnexinA2 expression in C4-2B cells is 0.22,in contrast with the internal reference,which was obviously lower than those of LNCaP and PC3 cells with lower metastasis potency(relative grey value is 0.93 and 0.95,respectively.P＜0.01).After RNAi was used in PC3 cells to down-regulate the expression of AnnexinA2,the growth became faster for PC3-ANXA2-siRNA cells than PC3,PC3-Lip and PC3-empty vector cells (P＜0.05).After RNAi was used in PC3 cells to down-regulate the expression of AnnexinA2,the ratio of apoptosis was detected by flowcytometry in PC3,PC3-Lip,PC3-empty vector and PC3-ANXA2-siRNA cells,and the apoptosis ratio in PC3-ANXA2-siRNA cells was the highest.However,the difference was not significant compare to others (P＞0.05).After RNAi was used in PC3 cells to downregulate the expression of AnnexinA2,the expression of AnnexinA2 in PC3-ANXA2-siRNA cells was decreased while the expression of MMP-2 and MMP-9 were increased.After RNAi was used in PC3 cells to down-regulate the expression of AnnexinA2,invasiveness of PC3-ANXA2-siRNA cells detected by transwell cabinet test was increased in vitro,and migration of PC3-ANXA2-siRNA cells detected by scratch test was increased in vitro.Conclusions The down-regulation of expression of AnnexinA2 could increase the invasive and metastatic ability of prostate cancer,and this may attribute to the up-regulation of MMP-2 and MMP-9 expression in prostate cancer.

Objective To evaluate GGT in combination with B ultrasound for the diagnosis of biliary atresia (BA) infants suffering from obstructive jaundice.Methods A retrospective analysis was made on 69 sick infants including 55 BAs and 14 non-BAs as identified by intraoperative cholangiography.The preoperative laboratory GGT and ultrasound data were collected and analyzed.The sensitivity,specificity,positive predictive value,negative predictive value and accuracy were compared.Results BA patients had significantly higher GGT than Non-BA patients (t =-4.164,P ＜ 0.05).The sensitivity,specificity,positive predictive value,negative predictive value and accuracy of GGT ＞ 306 U/L were 69.1％,92.9％,97.4％,43.3％,73.9％,respectively.In BA group,abnormal gallbladder was significantly associated with proadening portal vein,broadening hepatic artery compared with Non-BA patients (x2 =9.995,P ＜0.05).The accuracy of abnormal gallbladder on ultrasound was 78.3％.When two method combined for the diagnosis of BA,the sensitivity,specificity,positive predictive value,negative predictive value were 92.7％,92.9％,98.1％ and 76.5％ and accuracy can reach 92.8％.Conclusions For obstructive jaundice infants with GGT ＞ 306 U/L and abdominal gallbladder ultrasound finding intraoperative cholangiography should be carried out to make definite diagnosis of BA.

Objective To investigate the expression and significance of integrin αvβ8, p38 and extracellular signal-regulated protein kinase 1/2 (ERK1/2) proteins, which are TGF-β1 pathway related regulatory protein, in liver fibrosis of children with biliary atresia (BA). Methods Fifteen cases of BA (Kasai group) and 10 cases of congenital biliary dilatation (CBD group) were collected in Tianjin Children’s Hospital. And liver biopsy specimens were collected in Tianjin first central hospital, including 10 cases of BA children who underwent liver transplantation due to liver failure after Kasai operation (liver transplantation group). The specimens of front part of the right lobe of the liver were taken for HE and immunohistochemistry (IHC) staining. The expressions ofαvβ8, p38 and ERK1/2 in liver were observed by IHC staining in three groups of liver tissues. Results HE staining showed fibroblast hyperplasia occasionally in CBD group, portal area expansion, fibrous tissue proliferation and wide spread bridging fibrosis with few pseudo lobules in Kasai group. In transplantation group, portal area was widened, the degree of fibrosis was severe and bridging fibrosis generally formed resulted in pseudo lobules widely. Imunohistochemistry showed that the expressions of αvβ8 and ERK1/2 were weakly positive, and the expression of p38 was negative in CBD group. In Kasai group, the expressions of αvβ8, p38 and ERK1/2 proteins were all strongly positive in liver cytoplasm, biliary epithelial cells and vascular endothelial cell cytoplasm. In liver transplantation group the expressions of αvβ8, p38 and ERK1/2 proteins were all strongly positive. The semi-quantitative analysis showed that the expressions levels of αvβ8, p38 and ERK1/2 were significantly higher in Kasai and liver transplantation groups than those of CBD group (P<0.05). There were no significant differences in expression levels ofαvβ8, p38 and ERK1/2 between Kasai group and transplantation group (P>0.05). Conclusion The expressions ofαvβ8, p38 and ERK1/2 are gradually increased in liver of BA with the process of fibrosis, which indicate that they may be involved in
the process of BA liver fibrosis.

OBJECTIVE To investigate the distribution of clinical bacterial isolates and the change in antibiotic resistance spectrum in our hospital from 2005 to 2007.METHODS Data of bacterial susceptibility testing of clinical isolates from the Second Affiliated Hospital in of University of South China from 2005 to 2007 were collected and analyzed by software WHONET25.Results were assessed according to the National Committee for Clinical Laboratory Standards(NCCLS) of America issued in 2005.RESULTS The amount of Gram-negative bacteria decreased and of Gram-positive bacteria increased during this period.The proportion of coagulase negative Staphylococcus(CNS) had been increasing and reached 21.7% in 2007.The proportions of Staphylococcus aureus decreased from 17.6% in 2005 to 13.0% in 2007.Escherichia were the top two bacteria in 2007.The drug resistance rate of staphylococci against penicillin and erythromycin was more than 92.2% and 52.2%,respectively.The oxacillin resistance rate of CNS was 74.5%,significantly higher than that of S.aureus(16.5%).Drug resistance rate of Enterococcus to vancomycin was 1.1%.Gram-negative bacteria were found resistant to meropenem and imipenem.The resistance rate to ampicillin of Klebsiella and Escherichia was very high.CONCLUSIONS The variation of drug resistance and distribution of clinical bacterial isolates in our hospital are related to the improper use of antibiotics.It is very important to select antibiotics correctly according to the results of antibiotics susceptibility tests.

Epidermal growth factor receptor (EGFR) -mutant advanced non-small cell lung cancer (NSCLC) was previously regarded as a cold tumor according to tumor immune microenvironment (TIME) . However, recent studies have found that EGFR-tyrosine kinase inhibitors (EGFR-TKIs) treatment can transform the host immunity from immunosuppressive to immunosupportive state, bringing new hope for immunotherapy. There are four main therapeutic strategies for patients after EGFR-TKIs acquired resistance: immunotherapy alone (Im) , immunotherapy plus chemotherapy (Im+C) , immunotherapy plus antiangiogenic drugs (Im+A) , and immunotherapy combined with antiangiogenic drugs and chemotherapy (Im+A+C) . Among them, the efficacy of Im is extremely limited, being significantly lower than that of chemotherapy alone, while there is still scarce evidence for the efficacy of Im+A with few clinical studies. The combination of Im+C and Im+A+C shows better efficacy than chemotherapy alone. Im+A+C has a superior clinical outcome to Im+C. Additionally, the EGFR L858R mutation subgroup benefits more from Im+C than the EGFR 19 del mutation subgroup. The T790M-negative subgroup has a greater benefit from Im+A+C than the T790M-positive subgroup. In general, the strategy of combining immunotherapy with chemotherapy and/or an antiangiogenic drug represents a novel and promising method for treating EGFR-mutant NSCLC after EGFR-TKI failure.

Background and purpose:Hypermethylated in cancer 1 (HIC1) is silenced in multiple cancer cells and tissues by DNA methylation of epigenetic modification, which may modulate the initiation and progression of tumors. However, there are few reports about this phenomenon in prostate cancer. This study aimed to investigate the status of HIC1 promoter methylation in prostate cancer using methylation methods.Methods:Methylation-specific polymerase chain reaction (MSP) and bisulfate sequencing PCR (BSP) were used to detect the methylation status ofHIC1 promoter in prostate cancer cell lines PC3 and C4-2B, prostate normal cell line PrEC, primary Chinese PCa tissues and the respective healthy control cases.HIC1 expression level was respectively determined by reverse transcription-PCR (RT-PCR) and Western blot assays in PC3, C4-2B and PrEC cells treated with 5-Aza-CdR.Results:We found that the percentages of HIC1 promoter methylation were 78.23%, 72.15% and 10.63% in PC3, C4-2B and PrEC cells by MSP analyses. Moreover, the levels of methylatedHIC1 promoter in 36 primary Chinese PCa tissues compared with the respective healthy control cases were 80.30%vs 31.56%. Expressions ofHIC1 mRNA and protein level were restored in PC3 and C4-2B cells after 5-Aza-CdR treatment.Conclusion:These findings demonstrate thatHIC1 promoter region is hypermethylated in prostate cancer, which results in silence or downregulation ofHIC1. The status ofHIC1 methylation can be a valuable marker in the early stage of prostate cancer and a potential therapeutic target.

Background and purpose: Gene chip is a nucleic acid sequence analysis method which is based on hybridization. It is a high-through put assay which can widely detect the level of gene expression in different tissues and cell types. This study aimed to compare and bioinformatically analyze differentially expressed genes between higher malignant degree of prostate cancer tissues and prostate inflammation tissues. Methods: The total RNAs were isolated from tissues of prostate cancer and prostate inflammation by TRIzol method and then purified, reversely tran-scribed to cDNA with incorporating biotin labeling probe, hybridized with Affymetrix Human U133 Plus 2.0 (covering 47000 transcripts,representing 38500 distinct genes). Picture signals of fluorescence in gene array were scanned and differential expression of gene in two tissues were compared by Command Console Software 4.0. These differential expressed genes were analyzed by bioinformatics methods finally. Results: According to the fold change ≥2, P<0.05, 1819 differential expression genes including 1025 up-regulated genes and 794 down-regulated genes were discovered. GO enrichment analysis displayed that these differentially expressed genes were mainly involved in cell cycle, cell metabolism, etc. KEGG pathway analysis found that these genes were mainly involved in some metabolism pathways including purine nucleotide metabolism. The interactions between the proteins encoded by these genes were analyzed by STING. Twenty key nodes genes including TPX2, ANLN, NUSAP1, MELK, DLGAP5, KIF11, TOP2A, RRM2 were dis-covered. Then this study revealed CEP55 and ANLN might be related to the occurrence and metastasis of prostate cancer by looking through literature. Conclusion: During the development of prostate cancer, the activation of genes related to cell cycle and cell migration, the abnormalities of genes related to metabolism and the inhibition of genes related to cell adhesion play critical roles in the development of prostate cancer. CEP55 and ANLN were related to the occurrence and prognosis of prostate cancer by systematic analysis which provided a valuable clue for the next experiment.