Objective To explore the expression of annexin A10 (ANXA10) in human hepatocellular carcinoma (HCC),and analyze its correlation with matrix metalloproteinases-9 (MMP-9) and vascular endothelial growth factor (VEGF),thus to provide the basis for clinical diagnosis and assessment of HCC.Methods 88 HCC patients were selected,and they were all performed surgical treatment,HCC stage Ⅰ in 11 cases,stage Ⅱ 25 cases,stage Ⅲ 31 cases,stage ⅣV 21 cases.The expressions of ANXA10,MMP-9,VEGF of HCC cancer tissues,adjacent liver tissues and normal liver tissues were tested by immunohistochemical method.The correlation of ANXA10 with VEGF and MMP-9 was analyzed.Results The absorbance value of ANXA10 expression in the HCC cancer tissue was (0.074 ± 0.012),which was lower than that in the adjacent liver tissues [(0.091 ± 0.013)] and normal liver tissues[(0.131 ±0.025)],and ANXA10 expression of the adjacent liver tissue was lower than that of the normal liver tissues,the differences were statistically significant (t =8.96,9.44,8.71,all P ＜ 0.05).The absorbance values of MMP-9 and VEGF expression in the HCC cancer tissue were (0.147 ± 0.017) and (0.127 ± 0.028),respectively,which were higher than those in the adjacent liver tissues [(0.096 ± 0.012),(0.091 ± 0.015)] and normal liver tissues [(0.075 ± 0.014),(0.077 ± 0.019)].The absorbance values of MMP-9 and VEGF of the adjacent liver tissues were higher than those of the normal liver tissues,the differences were statistically significant (t =8.05,9.30,8.11;8.28,9.51,8.02,all P ＜ 0.05).The absorbance value of ANXA 10 expression in the HCC stage Ⅲ + Ⅳ cancer tissue was (0.056 ± 0.010),which was lower than that in the stage Ⅰ + Ⅱ cancer tissue [(0.082 ±0.016)],the difference was statistically significant (t =8.90,P ＜ 0.05).The absorbance values of MMP-9 and VEGF expression in the stage Ⅲ + Ⅳ cancer tissue were (0.157 ± 0.022) and (0.169 ± 0.033),respectively,which were higher than those in the stage Ⅰ + Ⅱ cancer tissue [(0.114 ±0.015),(0.091 ±0.021)],the differences were statistically significant (t =9.13,9.72,all P ＜ 0.05).ANXA10 was correlated with MMP-9 and VEGF (r =0.324,0.295,all P ＜ 0.05).Conclusion ANXA10 presents lower expression in HCC cell,and the expression decreased with the increase of staging.It is negatively related with the MMP-9 and VEGF.ANXA10 expression missing or inactivation of malignant change may be one of the most important features in HCC.

Indoleamine 2,3-dioxygenase 1 (IDO1) is the rate-limiting enzyme which catalyses the metabolism of L-tryptophan(L-Trp) in the kynurenine pathway.It is overexpressed in many tumor cells and antigen presenting cells.This enzyme inhibits local immune response and supports tumor cells to evade immune surveillance by depleting L-Trp and producing kynurenine metabolites,thus,it is an important target for cancer immunotherapy.There are several IDO1 inhibitors with different scarfold under investigation,three of which have already entered clinical stage.The role of IDO1 in tumor immune tolerance and the research progress on IDO1 inhibitors in recent years are summarized in this paper.

Type 2 diabetes,a glucose and lipid metabolism disorder accompanied by chronic multiple organ damage,has become a huge threat to human health,α/β hydrolase domain-6 (ABHD6) regulates the insulin release negatively by hydrolyzing monoacylglycerol.Small molecule ABHD6 inhibitors have been proven to lower bloodglucose and regulates energy homeostasis,which is a potential candidate for the treatment of type 2 diabetes.This paper introduced the ABHD6 signaling pathway and its mechanism,then reviewed the progress of small molecule ABHD6 inhibitors with different structures in recent years,and analyzed the structure activity relationship.

OBJECTIVE：To synthesize Tacrine,a new drug for the treatment of Alzheimer′s disease.METHOD:With the 2-aminobenzoic acid as the starting material，Tacring was synthesized via condensation-cyclization,chlorization and amination.RESULTS:Each step was modified and improved.The overall yields was 40.7%.The melting point and IR、1HNMR of the product prepared were identical as that of the reportedone.CONCLUSION:The process can be easily controled and is suitable for a scale production.

The effects of 4 kinds of edible oils on serum lipid levels, morphological changes of cardiovascular tissues, fatty acid compositions of various tissues and platelet function in rats had been observed. 40 adult Wistar rats fed semi-synthetic diet containing edible oil to supply 41% energy were divided into 5 group, i. e. control group (18.7% energy from fat), soy bean oil group, peanut oil group, lard group and rape seed oil group. The animals were fed the diets and water ad libitum for 2 months. The results showed that the lard gave the most serious detrimental effect but the soy bean oil was the least. The difference between these two groups was significant. The platelet number was 223?109/L and aggregation rate was 21.8% of the soy bean oil group but. the lard group 149?109/L and 30.2%. The composition (%) of PUFA in tissues was higher and that of saturated fatty acid was tower in soy bean ou group than those in lard group.Electron microscopical studies showed that the animals with vascular endothelial cells changes were more and the pathological changes were more serious in the lard group than those in the soy bean group.All groups except rape seed oil group increased weights in the experi-mental period at the similar rate. Rape seed oil group gained very little weights and aggregated more C22:1 especially in myocardium.

Objective To screen the mutations of MYOC gene in a Chinese primary open angle glaucoma (POAG) family from Cbengqing and investigate the relationship between the mutations in MYOC/TIGR gene and POAG.Methods In a large 4-generation glaucoma family, myocilin gene (MYOC) was screened in 39 family members, 8 of which were confirmed patients. Normal controls included 100 normal Chinese subjects.The known mutations of MYOC gene ( including G34C, C136T, G144T, G227A, C624G,G736A, C1009G, A1036G, C1081T, G1099A, G1138A, A1139C, T1430A, C1441A and C1442T) were detected by single strand conformation polymorphism(SSCP) , po]ymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis and DNA sequencing.Results G227A mutation was detected in 2 POAG patients and 1 asymptomatic patient, but not in the controls.Cl009del mutation was identified in all patients of the pedigree and an offspring member but not in the controls. No other mutations were detected.Since the C1009del mutation was revealed for the first time, a new GenBank number FJ237047 correponding to ACI62293 was applied.Conclusions The G227A mutation is a known site and there is no relationship between G227A mutation and glaucoma. But C1009del may be related to glaucoma which suggests that morbidity could be higher in the relatives of POAG than the controls.

Objective To analyze the differential gene expression profiling of liver in rats subjected to hemorrhagic shock(HS) and sham hemorrhage shock(SHAM) by gene chip technology, thus to evaluate the possible molecular pathogenesis of HS. Method 20 male Wistar rats were randomly divided into a SHAM group and a HS group, with 10 rats in each group. Hepatic gene expression profiles were detected by oligonucleotide microarrays of 5705 mouse genes in two groups for three times. Genes with ratio(R) > 2 were identified as up-regulated and R < 0.5 were identified as down-regulated. Biological function of differentially expressed genes was analyzed and 9 genes were selected to undergo semi-quantitative RT-PCR. Results Among the total 5705 probes detected,86 genes showed differential expression in HS group comparison with SHAM group. The expression levels of 72 genes were up-regulated while those of 14 genes were down-regulated significantly. Differentially expressed genes were classified according to their biological function: transport genes, transcription regulator genes, signaling genes, response to stress genes, metabolic genes, development genes and cell adhesion genes. Conclusions cDNA microarray is an efficient and high-throughout method to survey gene expression profiles in HS.The variation of those gene expressions might be a potential pathogenic mechanism for HS that may offer a novel target for further study of therapeutic strategies of HS.

Through comparative study on Naotaifang containing serum and plasma proteomics (peptide), this article revealed differential proteins (peptides) in the Naotaifang. The characteristics of differential proteins were identified with mass spectrometry. It provides scientific evidences for the pharmacodynamic material basis and Chinese herbal medicine plasma pharmacological method development in the exploration of Naotaifang. A total of 20 healthy adult SD rats were randomly divided into the control group, Naotaifang treatment group according to their weights. Ten rats in each group. Intragastric administration of medication was given for seven consecutive days. Before surgery, rats were fed with water but without food. One hour after the last drug administration, 10% chloral hydrate was injected for intraperitoneal anesthesia. Blood was taken through the common carotid artery. Serum and plasma samples were made after blood was taken from each rat. Serum and plasma samples of five rats were randomly selected from each group. And the two-dimensional electrophoresis (2-DE) technique was used in the comparative study of serum pro-teomics (peptide). The 300 DPI scanning and PDQuest 7.3.0 were used in the analysis. The ESI-MS/MS was used to identify important differences in proteins and screen characteristic serum and plasma protein. The results showed that 20 differential proteins of 5 plasma samples were identified. There were 15 types of proteins expressing up-regulation and 5 types expressing down-regulation. Comparative analysis on the 2-DE gel pictures of Naotaifang containing serum, 19 differential proteins of 5 plasma samples were identified, among which 15 types of proteins express up-regulation and 4 down-regulation. Comparative analysis on the 2-DE gel pictures of Naotaifang containing serum and Naotaifang containing plasma showed that 24 differential proteins of 5 plasma samples were identified, among which 9 types of proteins express up-regulation and 15 down-regulation. The highly expressed proteins were selected to MALDI-TOF-MS between Naotaifang containing serum and Naotaifang containing plasma. There were six successful-ly identified proteins, which were inter-alpha trypsin inhibitor, heavy chain 3, group-specific component, comple-ment factor B, Receptor Complexed with A Heterodimeric Fc, isoform CRA-d, Transferrin. It was concluded that protein with obvious changes in the Naotaifang containing serum and plasma may be related with fibrinolysis and an-ticoagulant. These proteins are involved in angiogenesis, inflammation and other pathological regulations of physiolog-ical processes. They are of great significance in the study of effective target and its signal transduction pathway of Naotaifang.

Objective To estimate the influence of palmitic acid (PA) on the proliferation of and production of inflammatory mediators by a human keratinocyte line HaCaT.Methods Cultured HaCaT cells were treated with PA of eight concentrations (0-200 μmol/L) for 3-24 hours followed by the evaluation of cell proliferation by using the cell counting kit-8.According to the proliferation assay,four concentrations (75,100,125,150 μmol/L) of PA were selected and used to treat HaCaT cells for 24 hours,then,fluorescence-based immunohistochemical staining was performed to observe the nuclear translocation of nuclear factor (NF)-κB p65,enzyme linked immunosorbent assay (ELISA) to determine the level of interleukin (IL)-6 in the supernatant of culture medium,real-time PCR to detect the mRNA expressions of peroxisome proliferator-activated receptor oα (PPARα) and IL-6,and Western blot to quantify the protein expressions of PPARα as well as total and nuclear NF-κB p65.Those HaCaT cells receiving no treatment served as the control group.Statistical analysis was carried out by one-factor analysis of variance using the GraphPad Prism 5.0 software.Results The HaCaT cells treated with PA of 50-175 μ mol/L showed accelerated proliferation compared with the control HaCaT cells (all P ＜ 0.05).PA from 75 to 150 μmol/L enhanced the nuclear translocation of NF-κB p65,mRNA and protein expressions of PPARα,as well as the mRNA expression and supernatant level of IL-6 in a dose-dependent manner.The relative expression level of nuclear NF-κB p65 protein was 0.4536 ± 0.0173,0.5184 ± 0.0206,0.5333 ± 0.0231,0.6160 ± 0.0297,and the supernatant level of IL-6 was (31.5677 ± 0.2268),(32.3773 ± 0.4156),(32.9837 ± 0.0029) and (33.6890 ± 0.0936) ng/L,in HaCaT cells treated with PA of 75,100,125 and 150 μmol/L,respectively,compared to 0.3237 ± 0.0114 (all P ＜ 0.01) and (30.4577 ± 0.5131) ng/L (all P ＜ 0.01) in the control HaCaT cells,respectively.Conclusions PA can accelerate the proliferation of HaCaT cells,enhance NF-κB nuclear transfer,PPARα expression and IL-6 secretion in a dose-dependent manner within a certain concentration range,and may exert a promoting role in the activation and expression of some inflammatory factors.

Objective To observe the effect of sirolimus,an autophagy enhancer,on premature senescence in fibroblasts induced by repeated exposure to a subtoxic dose of ultraviolet B (UVB).Methods Skin fibroblasts from foreskin tissue of healthy adolescents were classified into six groups:control group cultured in Dulbecco's modified Eagles' medium (DMEM) containing 1％ calf serum,UVB group receiving UVB irradiation only,sirolimus group treated with sirolimus of 10 mg/L (added after daily exchange of culture medium),and three combined groups receiving UVB irradiation immediately followed by overnight treatment with sirolimus of 0.1,1.0 and 10.0 mg/L respectively.UVB irradiation was given at a dose of 10 mJ/cm2 once a day for five successive days.After five days of treatment,cell counting kit-8 (CCK-8) was used to evaluate cell viability,β-galactosidase staining to detect senescent ceils,Western blot to quantify the expressions of p53,LC3-B and beclin 1 in these fibroblasts.Autophagy level was determined by acridine orange staining followed by fluorescence microscopy and transmission electron microscopy.Data were processed by the SPSS 16.0 software,and statistical analysis was done by one-way analysis of variance,t test and least significance difference.Results Sirolimus significantly increased the proliferative activity of fibroblasts in a dose-dependent manner,with the absorbance value at 450 nm being 0.27 ± 0.02,0.36 ± 0.04 and 0.39 ± 0.04 for fibroblasts irradiated with UVB followed by treatment with sirolimus of 0.1,1.0 and 10 mg/L respectively,compared to 0.26 + 0.01 for fibroblasts irradiated with UVB only (all P ＜ 0.05).Significant differences were also observed between the fibroblasts irradiated with UVB followed by treatment with sirolimus of 0.1,1.0 and 10 mg/L and those irradiated with UVB only in the percentage of β-galactosidasepositive fibroblasts (92.50％ ± 0.34％,42.40％ ± 0.53％ and 6.20％ ± 0.39％ vs.95.10％ ± 0.32％,all P ＜ 0.05)and intracellular intensity of acridine orange-induced fluorescence (36.43 ± 0.24,45.25 ± 0.33 and 48.69 ± 0.37 vs.33.99 ± 0.32,all P ＜ 0.05).Moreover,the expressions of p53,LC3-B and beclin 1 in the three combined groups differed significantly from those in the UVB group (all P ＜ 0.05).Conclusion Sirolimus can inhibit UVBinduced premature senescence likely via upregulation of autophagy in fibroblasts.