OBJECTIVETo study the inhibitory effect of 10-gingerol on the proliferation of hepatocellular carcinoma HepG2 cells and the role of Src/STAT3 signaling pathway in mediating the effect.

METHODSSYBYL-X2.1 software was used to simulate the interaction between 10-gingerol and Src. HepG2 cells treated with 10-gingerol at 1, 3, 10 or μol/L for 24 h were assessed for cell viability using MTT assay, and EdU staining was used to detect the cell proliferation and calculate the number of positive cells. The expressions of p-Src and p-STAT3 were detected using Western blotting, and the mRNA expressions of the target genes of STAT3 (cyclin D1 and CMCC) were detected using qPCR.

RESULTS10-gingerol was capable of forming hydrogen bond with such Src residues as TRY-340, MET-341, MET-314, ASP-404, and ILE-336. MTT assay showed that 10-gingerol at 3 and 10 μmol/L significantly lowered the viability of HepG2 cells ( < 0.001). Treatment with 1, 3, and 10 μmol/L 10-gingerol significantly reduces the number of EdU-positive HepG 2 cells ( < 0.001). Western blotting showed that 10-gingerol at 3 and 10 μmol/L significantly decreased the phosphorylation levels of Src and STAT3 in HepG2 cells ( < 0.01). 10-gingerol at 1, 3, and 10 μmol/L significantly decreased the mRNA expressions of cyclin D1 and CMCC as shown by qPCR ( < 0.01).

CONCLUSIONS10-gingerol can dose-dependently inhibit the proliferation of HepG2 cells and suppress the activation of Src and STAT3.

Objective To investigate the mutations of IDH1,IDH2,p53 gene,and Ki-67 protein expression in different grade of gliomas and identify the association with its clinical relevance. Methods The mutations of IDH1,IDH2 and p53 gene were detected by direct DNA sequencing,and protein expression of Ki-67 was analyzed by immunohistochemistry. The correlations between gender,age,tumor site,differentiation degree and pathological type of patients were analyzed. Results R132H mutation of IDH1 gene was detected in 32.6% samples (14/46 cases),of which the proportion of WHO classification grade Ⅱ was 40.0%,and grade Ⅲ was 58.3%. IDH1 mutations were shown correlated with age,pathology level Ⅱ-Ⅲ,and Ki-67 low expression. p53 mutations were detected in 4 glioblastomas,with mutations located at exon 7,8. IDH1 gene mutation was negatively correlated with Ki-67 expression. Conclusions The proportion of IDH1 gene mutation in different pathological types of gliomas is different,which is the highest in gradeⅡ~Ⅲ. It is suggested that the subtypes should be listed independently by routine tests. Mutations in p53 gene are more common in primary glioblastomas and may be associated with adverse outcomes. The combined detection of DH1,p53 and Ki-67 is conducive to the diagnosis and prognosis of glioma.

Antiviral Agents ; Diabetic Retinopathy ; Drug Combinations ; Hepatitis C, Chronic ; Humans ; Interferon-alpha ; Polyethylene Glycols ; Recombinant Proteins ; Ribavirin

Objective To observe the influence of Buyang Huanwu Decoction ( BYHWD) on the inhibition of rat myocardial H9C2 cell activity and SH2-containing tyrosine phosphatase-1 ( SHP-1) activity induced by trastuzumab, and to explore the possible regulatory mechanism after observing the intervention of BYHWD on rat myocardial H9C2 cell transfected with SHP-1 or SHPC/S-1 gene. Methods The eukaryotic expression vectors pcDNA3.1 (+)- SHP-1 and pcDNA3.1 (+) -SHPC/S–1 were constructed and then were transfected to rat myocardial H9C2 cells using the method of liposome transfection. The cells with positive clones were screened out with G418, and then were cultured with trastuzumab for maintaining growth. Using quantitative RT-PCR, we detected the expression of SHP-1 gene and SHPC/S - 1 gene in rat myocardial H9C2 cells. The phosphatase activity analysis was used for observing the regulatory effect of BYHWD on SHP-1 in myocardial cells. Furthermore, we observed the apoptosis of rat myocardial H9C2 cells by methyl thiazolyl tetrazolium (MTT) assay after treatment with BYHWD. Results Sequencing results indicated the successful construction of eukaryotic expression vectors, which had stable expression in myocardial H9C2 cells even under the intervention of trastuzumab. The results of phosphatase analysis showed that H9C2-SHP-1 had the highest activities of phosphatase, but the activities were decreased after the intervention with BYHWD ( P<0.05) . The results of MTT assay also showed the apoptotic rate of H9C2-SHP-1 cells was decreased after treatment with BYHWD ( P <0.05) . Conclusion BYHWD can promote the proliferation of myocardial H9C2 cells inhibited by trastuzumab, and can regulate the expression of SHP-1 in myocardial cells, which will supply reference to the further study of treatment of trastuzumab-induced cardiac toxicity.

OBJECTIVETo study the potential factors in influencing the performance of immunohistochemical testing for HER2 protein in gastric cancers.

METHODSThe HER2 protein expression status of 1 471 surgically resected archival gastric cancer cases in Drum Tower Hospital collected during two different periods was retrospectively analyzed. The materials included 957 cases tested during the period from 2007 to 2009 (group 1) and 514 cases from 2012 to 2013 (group 2). The test procedures and results observed during these two periods were compared.

RESULTSThe percentages of score 3 HER2 protein expression (14.4%, 74/514 versus 9.5%, 91/957) and score 2 or score 3 HER2 protein expression (27.2%, 140/514 versus 21.7%, 208/957) were both higher in group 2 than in group 1 (P < 0.05). In group 1, the cancer tissue was fixed in 10% formalin, stained manually with HER2 antibody A0485 (Dako) and assessed by different pathologists.In group 2, the tissue was fixed in 10% neutral buffered formalin (pH 7.2), stained using automated immunostaining system (Roche Benchmark XT) with HER2 antibody 4B5 (Ventana) and assessed by a specialized team of pathologists.

CONCLUSIONThe results of HER2 immunostaining in gastric cancer are influenced by a number of factors including type of fixative, clone number of primary antibody, staining methods and experience of pathologists.

Antibodies, Monoclonal ; Fixatives ; Formaldehyde ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Receptor, ErbB-2 ; metabolism ; Retrospective Studies ; Staining and Labeling ; Stomach Neoplasms ; metabolism

Seventy patients with advanced non-small-cell lung cancer (NSCLC) aged 65 or above were treated with epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) erlotinib or gefitinib from February 2006 to September 2010. The efficacy and toxicities of treatment were retrospectively analyzed.The overall response rate and disease control rate were 31.4％ and 84.3％,respectively. Themedian progression-free survival time and median survival time were 8.0 months and 13.5 months,respectively(P ＜ 0.05 ). One-year survival rate was 54.3％. Response rate ( CR + PR) ( 42.9％ ) anddisease control rate (94.3％ )in female patients were superior to males (20.0％ and 74.3％ ) (P ＜ 0.05 ).Non-smoking and PS score ＜ 2 were good predictors for survival.The side effects were generally mild and mainly were skin rash and diarrhea.

Objective To observe the expression of Her-2/neu protein and androgen receptor (AR) in human prostate cancer and to evaluate their significances in the progression of prostate cancer. Methods The Her-2/neu protein and AR immunohistochemical stain were carried out in human prostate tissue microarray that consisted of prostate cancer (107 cases) and benign prostate tissue (42 cases). The prostate cancer cases were divided into 4 groups: group one (Gleason score 6),group two (Gleasonscore 7), group three (Gleasonscore 8) and group four (Gleasonscore 9) according to the Gleason score. The immunostains immunohistochemical stain were interpreted in two aspects of the staining intensity and the percentage of positive cells. The significance and relationships between the expression of Her-2/neu protein and AR in prostate cancer and benign prostate tissue (BPT) and the grouping of different Gleason scores of prostate cancer were then evaluated. Results The positive expression rate of Her-2/neu protein was significantly higher in prostate cancer tissue than in BPT [43.9%(47/107) vs. 14.3%(6/42), x2=11.562, P=0.009], and the positive expression intensity of Her-2/neu immunoreactivity was also higher (x2= 11.764, P=0.008). There were significant differences in positive expression intensity of Her-2/neu immunoreactivity among the different Gleason scores groups (x2 = 20. 512, P = 0. 015), and the expression intensity was significantly positively correlated with Gleason scores ( r= 0. 269, P = 0. 005). There was significant difference in AR immunoreactivity between in prostate cancer (67 %, 72/107) and in BPT (50 %, 21/42, x2 =3. 843, P=0. 050). Among prostate cancer cases, the positive expression intensity of AR was not significantly different among groups 1 through 4 (x2 = 4. 318, P = 0. 229), and was not significantly correlated with Gleason scores ( r = - 0. 065, P = 0. 505 ). Moreover, the positive expression intensity of Her-2/neu protein was not significantly correlated with that of AR (r = -0. 115, P=0. 237). Conclusions Overexpression of Her-2/neu protein in human prostate cancer tissue suggests that Her-2/neu may have some role in prostate tumorigenesis. Her-2/neu protein expression is positively correlated with Gleason score in prostate cancer, which suggests that Her-2/neu may be a potential prognostic predictor of prostate cancer.

Objective To investigate the inhibitory mechanism of emodin on lipopolysaccharide(LPS)-induced nitric oxide(NO)generation in rat peritoneal macrophages.Methods NO production and iNOS expression were measured through nitrite assay and Western blotting assay,respectively.NF-kB activity and nuclei P65 expression were estimated by dual-luciferase and Western blotting assay,respectively.Intracellular free Ca2+([Ca2+]i)was detected using the ratiometric fluorescent calcium indicator dye,Fura-2,and a microspectrofluorometer.PLC-γphosporylation was analyzed by Western blotting assay.Results First,emodin was found playing active roles in suppressing LPS-induced NF-kB activation in rat peritoneal macrophages.Second,emodin down-regulated transient[Ca2*]i and could increase in NF-kB upstream signal.Finally,emodin suppressed phosphorylation of PLC-γ by LPS stimulation in the upstream of[Ca2+]i.Conclusion Suppression of PLC-γ phosphorylation is involved in emodin inhibiting NO generation by LPS stimulation in rat peritoneal macrophages.

Objective To evaluate the expression of molecular markers in prostate cancer and to clarify the significance of these markers as prognostic indicators for androgen deprivation therapy. Methods A series of 116 prostate cancer patients under androgen deprivation therapy as a single treatment was reviewed. Expression levels of 7 proteins, including androgen receptor(AR),E-cadherin, Chromogranin A(CgA) , Ki-67, Survivin, EZH2 and hepsin, were measured by immunohistochemical staining. Results Of the 7 molecules. Ki67,EZH2 and Survivin expression were significantly associated with several conventional prognostic factors. Univariate analysis identified clinical stage, Glea-son scores,pretreatment serum PSA level, Ki-67 and Survivin expression as significant predictors for prostate-specific antigen (PSA) progression after endocrine therapy. Of these significant factors, Survivin expression, clinical stage and Gleason scores appeared to be independently related to PSA progression after endocrine therapy by multivariate analysis. Furthermore, there were significant differences in PSA progression-free survival according to positive numbers of these three independent risk factors. Conclusion Survivin could be a useful independent prognostic factor in prostate cancer with endocrine therapy, besides clinical stage and Gleason score.

Objective To explore the relationship between polymorphism of G-protein ?3 subunit (GNB3) gene and the emotion trait of chest stuffiness and pains patient. Methods The GNB3 gene type in 27 patients of depression of chest stuffiness and pains, 40 patients of no depression of chest stuffiness and pains, and 20 patients of healthy subjects (control) was determined by polymerase chain reaction and restriction fragment incision enzyme, and score of depression was evaluated. Result Occurrence of depression in chest stuffiness and pains was significantly higher than that in control (P