Objective To study the expression-mode and dynamic transmutation of high mobility group box-1 (HMGB1) in hepatocytes of the mouse model with acute hepatic failure and to study the interaction beween HMGB1 and tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β). Methods The mouse model of acute hepatic failure was established by injecting D-galactosamine (D-GalN) and lipopolysaccharide(LPS). Immunohistochemistry SABC method was used to detect the HMGB1 expression at 6 time points. The enzyme linked immunosorbent assay (ELISA) was used to determine serum TNF-α. IL-1β levels. Paired t test was used for statistical analysis. Results The HMGB1 expression was detectable at 2 hours after injection, which dramatically increased over time and peaked at 24 hours after injection. The serum TNF-a level and IL-1β level increased right after injection. The TNF-a level peaked at 8 hours after injection with a maximum value of (473.42±22. 99) pg/mL. The IL-1β level peaked at 2 hours after injection with a maximum value of (724. 49±34. 24) pg/mL. Both cytokine levels slowly decreased after peaking. IL-1β level returned normal with (51. 49±36. 87) pg/mL. Conclusions HMGB1 is one of the most important factors during the development of acute hepatic failure, which can promote the secretion of TNF-α and IL-1β at early stage and be abundantly expressed under the effect of these cytokines at middle and late stages with the result of liver damage. This process is directly correlated with the development and severity of hepatic failure.

Objective:To evaluate the clinical sensitivity and specificity of sentinel lymph node biopsy (SLNB) for oral cancer.Methods:By searching Medline,CENTRAL,EMBSE,CBM,CNKI,WANFANG,etc,an studies about SLNB for the patients with oral cancer array for lymph node metastasis were retrieved.The quality of included studies was evaluated by QUADAS items.The data were analyzed by statistic software Meta-DiSc 1.4.Results:25 studies including 934 subjects were analyzed in the current systematic review.The pooled values of sensitivity,specificity were 91％ (95％ CI 88％-94％) and 100％ (95 ％ CI 99％-100％) respectively.The area under summary receiver operating characteristic (SROC) curve (AUC) of SLNB for cervical lymph node metastasis was 0.99.Conclusion:SLNB procedure has high sensitivity and specificity in the diagnosis of cervical lymph node metastasis,it can be used as an useful method for cervical lymph node metastasis screening in patients with oral cancer.

To investigate peripheral blood mononuclear cells (PBMCs) infected by hepatitis C virus (HCV) and the influenceon T lymphocyte subset. Methods: Using non-isotopic in situ hybridization (NISH) and streptadivin-biotin-enzyme complex assay (SABC) todetect HCV-RNA, NS5 antigen and T lymphocyte subset. Results: 8(40.0% ) out of 20 patients with hepatitis C virus were HCV-RNA posi-tive in PBMCs, and among them 6(30.0％) were NS5 antigen positive simultaneously. NS5 antigen signals appeared diffusely within cyto-plasm, or in cell membrane, while HCV-RNA signals were mainly distributed in cytoplasm. Both the proportion of NS5 positive cell and HCV-RNA positive cells were very low (≤3％).The proportion of positive cellsane, while HCV-RNA signals are very low (≤3％). The proportionof CD4+ T cells was lower in the patients comparing with that in control group, the proportion of CD8+ T cells was higher, and the ratio ofCD4+/CD8+ were decreased remarkably (P＜0.01). Moreover the proportion of CD4+ T cells was lower in HCV-RNA-positive group com-paring with that in HCV-RNA-negative group, the proportion of CD8+ T cells was higher, and the ratio of CD4+/CD8+ was reversed (P＜0.01). Conclusion:HCV can infect PBMCs and replicate in patients with chronic hepatitis C;CD4+ T cells are readily damaged after PBMCsinfected by HCV, which causes the decrease or reversal of the ratio of CD4+/CD8+, undermines the ability of the body clearing virus and re-sults im chronicity of HCV infection.

Objective To investigate the effect of the balance between helper T lymphocyte (Th) 1 cytokines and Th2 cytokines in the serum of hepatic fibrosis rats induced by CCl4 on the expression of β-catenin in their livers. Methods Seventy-five rats were divided into control group,treatment group and model group. Hepatic fibrosis models were built by subcutaneous injection of CCl4in male Wistar rats, then rats in treatment group were treated with pentoxifylline(PTX) by intragastric administration. The model rats without PTX treatment served as treatment control. The serum levels of Th1 cytokines(interferon-γ, IFN-γ; tumor necrosis factor-α, TNF-α), Th2 cytokines (interleukin-4,IL-4;interleukin-6,IL-6) and type Ⅳ collagen were measured by enzyme-linked immunosorbent assay (ELISA). The expression and distribution of β-catenin in liver tissue and the percentage of area of type Ⅳ collagen accounted in the liver were analyzed by immunohistochemical analyses. The changes in serum cytokines were depicted by using One-way ANOVA, and the correlation between β-catenin expression and cytokines levels or hepatic fibrosis development was elucidated by Bivariate.Results The serum levels of Th2 cytokines in model group were significantly higher than those in treatment group, and even higher than those in control group (F=71. 260,91. 732, all P＜0.05).However, the levels of Th1 cytokines in model group were significantly lower than in treatment group while even lower than those in control group(F= 50. 420,10. 625, all P＜0.05). Serum cytokines came out to be Th2 polarized. Meanwhile, the percentage of type Ⅳ collagen area accounted in the liver,which was corresponding to the degree of liver fibrosis, was lower in the treatment group than the model group and even lower in control group (F=3 000,both P<0.05). β-catenin expression in the liver of model group was stronger than that in the treatment group, and even stronger than that in normal group (F=92. 030, both P＜0. 05). According to β-catenin immunohistochemical analyses based on patho-photos, livers from the model group showed dark color in both cell cytoplasm and nucleus, while the liver from the treatment group only showed weak color in cytoplasm and no straining color in nucleus and the liver from normal group showed colors neither in cytoplasm nor in the nucleus.The expression of β-eatenin had a positive correlation with either the level of Th2 cytokines in serum of rats (r=0. 560,P＜0.01) or the percentage of area of type Ⅳ collagen accounted in the liver of rats (r=0. 757, P＜0. 01). Conclusions Th2 polarization in serum cytokines can aggravate the development of liver fibrosis by inducing the expression of β-catenin, which in the livers of rats is a reflection of the degree of the liver fibrosis.

Objective: To study the influence of HMGB1-Abox on the expression of HMGB-1 in RAW264.7 cells stimulated by lipopolysaccharide(LPS)and its inhibition on the secretion of inflammatory factor.Methods: Use cloning vector to make up prokaryotic plasmid PET28a-HMGB1-Abox, which included the gene fragment of the A box of HMGB1, and recombinant vector was further transformed into Escherichia coil strain BL21(DE3),the recombinant plasmid was induced by isopropylthiogalactoside(IPTG)to express target protein.The protein was purified by Ni-column.We tested the effect of the different concentration of rHMGB1-Abox on the viabihty of RAW264.7 cells stimulated by LPS using CCK-8.Experimental group(EG)was treated with rHMGB41-Abox and control group(CG)was deh with PBS.TNF-α and IL-1β levels were detected by the enzyme linked immunosorbent assay(ELISA).Western blot and immunofluorescence staining method were used to compare the expression of HMGB1 in RAW264.7 cells with the experiment group and control group.Reverse transcription polymerase chain reaction(RT-PCR)was used to detect the variation tendency of HMGB1-mRNA.All procedures were manipulated at 2 \ 6 \ 12 \ 24 \ 48 h after treatment.Results: The recombinant prokaryotic expression vector PET28a-HMGB1-Abox was successfully constructed and mouse HMGB1-Abox protein about 14 kD was successfully expressed.There was a positive correlation between the viability of RAW264.7 cells and the concentration of protein and acting time.Compared to CG, TNF-α and IL-1β levels in EG declined significantly.In EG, Western blot and Immunofluorescence staining showed that the expression of HMGB1 protein was decreased and the expression of HMGB1-mRNA was reduced markedly and delayed.Conclusion: The rHMGB1-Abox could reduce expression and secretion of HMGB1 in RAW264.7 cells stimulated by LPS significantly,thereby to prevented the promotion of HMGB1 on pro-inflammatory mediator and inhibit the expression and secretion of inflammatory factors,which has some anti-inflammatory action.

Objective:To evaluate the clinical value of 64-slice spiral coronary CT angiography(CTA)in diagnosis of in-stent restenosis(ISR)after percutaneous coronary intervention(PCI)in patients with coronary artery disease.Methods:CTA was used to reconstruct and analyze the 345 segments(each stent was divided into three segments:proximal,middle and distal)of 115 stents in 60 patients with coronary heart disease after PCI.The results of selective coronary angiography(SCA)were taken as the golden standard to evaluate the sensitivity and specificity of CTA in diagnosis of ISR after PCI.Results:CTA clearly showed the location and length of the stents,the stenosis at stent and the characteristics of restenosis plaques.Thirty-nine in-stent restenosis lesions were found by CTA,including 8 calcified lesions and 31 non-calcified lesions.There were 25 lesions at the proximal end of the stent,7 at the middle and 7 at the distal end.Forty-two lesions were found by CTA,including 9 calcified lesions and 33 non-calcified lesions,with 26 at the proximal end,8 at the middle and 8 at the distal end.CTA correctly diagnosed 36 segments,missed 6,and misdiagnosed 3.The sensitivity,specificity,PPV,NPV and accuracy of CTA were 85.71%,99.01%,92.31%,98.04% and 97.39%,respectively.CTA had the highest sensitivity and specificity for diagnosis of the proximal ISR,being 96.15% and 100%,respectively.Proximal stent restenosis accounted for 64.10% of the total.Conclusion:CTA can clearly demonstrate the in-stent restenosis and has a high accuracy in diagnosing restenosis after PCI.CTA is a safe,simple and reliable noninvasive diagnostic method for diagnosis of in-stent restenosis after PCI.

OBJECTIVE To investigate the effects of somatostatin(SST)on expression of NO,TNF? and 5-lipoxygenase(5-LO)in primary cultured rat Kupffer cells induced by LPS.METHODS Kupffer cells isolated from Sprague-Dawley rats were cultured in the presence of LPS added alone or with SST for 12,24 and 48 h,and the production of TNF? was assessed in culture supernatants by ELISA and that of nitric oxide(NO)by nitrate reductase method.5-LO mRNA expression was assessed by semiquantitative RT-PCR.RESULTS Vehicle-stimulated Kupffer cells produced a basal amount of NO,TNF? and expressed 5-LO mRNA.Kupffer cells stimulated by LPS secreted significantly increased amounts of NO and TNF?(P0.05).CONCLUSIONS SST modulates NO,TNF? production and 5-LO mRNA expression by LPS-stlimulated Kupffer cells.

Objective To investigate the image quality and diagnostic accuracy using 64-slice spiral computed tomography (64-CTA) scanner in patients with suspected coronary artery disease. Methods Sixty eight patients with chest pain or post PTCA underwent CT coronary angiography (CTA) and selected coronary angiography (SCA). The SCA results were served as "gold standard" to evaluate the diagnostic accuracy of CTA, while the sensitivity, positive predictive value (PPV) and negative predictive value (NPV) were calculated, respectively. Results 64-slice spiral CT could clearly demonstrate the coronary arterial trunk and branchs with stenosis, calcifications abnormal orifise origination and bridge vascular disease; especially with high accuracy in revealing calcification and even with quantification. The sensitivity, specificity, PPV and NPV of the degree of stenosis more than 75% for coronary artery segments evaluated by CTA were significantly higher than those of the degree of stenosis less than 50% for coronary artery segments(P

0 05), respectively The overall rates of post-operative complications in the two groups were 11 5% and 24 7%(P

A mouse model for acute hepatitis B virus (HBV) infection was established by using the hydrodynamical injection of mouse tail vein, in which the immunocompetent BALB/c mice were hydrodynamically injected with a competent replication plasmid pAAV-HBV1.2 having 1.2 fold over-length of HBV DNA. On day 1, 2, 4, 6 and 8 after injection, the levels of HBsAg, HBeAg and HBV DNA in blood serum were detected by using ELISA and fluorogenic quantitative PCR assay (FQ-PCR). And on day 8. HBsAg and HBeAg in liver tissue were assayed by immunohistochemical staining. It was found that HBsAg in blood serum could be detected on day 1 after infection in 14 of 16 mice (85.7%) injected with pAVV-HBV1.2 by using ELISA assay and the peak levels of HBsAg and HBeAg were attained during the first day after injection and then it dropped down gradually up to day 8 following injection. The titer of HBV DNA in blood serum attained its peak on day 2 and maintained a high level later on. On day 8 after injection, its titer was 1.9×10~4 copies/mL. The percentage of HBcAg-positive hepatocytes and HBsAg-positive hepatocytes in liver tissues were 5% and 2% respectively. Thus, by using the hydrodynamic injection with the competent replication plasmid, a mouse model for acute HBV infection is successively developed.