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Objective: To analyze the short-term and long-term effectiveness of radiofrequency ablation (RFA) combined with transcatheter arterial chemoembolization (TACE) in the treatment of hepatocellular carcinoma (HCC)., Methods: The clinical data of 114 HCC patients, 104 males and 10 females, aged 55 (30 - 81), treated by RFA combined with TACE and followed up for 20 (1 - 82) months were analyzed., Results: Complete necrosis was achieved in 101 patients (88.6%). 10 patients showed incomplete necrosis and 3 patients showed new neoplasm, and they all underwent repeated RFA or TACE. No therapy-relative death was found. The overall 1-, 2-, 3-, 4-, and 5-year survival rates were 90.4%, 82.6%, 73.2%, 63.5%, and 49.1%respectively. The 1-, 2-, 3-, 4-, and 5-year tumor progression-free survival rates were 77.1%, 64.6%, 54.6%, 46.8%, and 36.4% respectively. The overall 1-, 2-, 3-, 4-, and 5-year survival rates for the tumors with the size < or = 5 cm and the tumors with the size of 5.1 - 7 cm were 95.5%, 84.6%, 73.1%, 61.5%, and 50.6% and 80.2%, 64.9%, 56.3%, 45.3%, and 39.5% respectively (P = 0.041). The overall 1-, 2-, 3-, 4-, and 5-year survival rates for the solitary tumor and multiple tumors (no more than 3 tumors) were 95.8%, 89.1%, 78.1%, 67.1%, and 56.7% and 80.0%, 60.6%, 46.6%, 33.4%, and 21.5% respectively (P = 0.001). The levels of albumin and alpha-fetoprotein, and boundary and number of tumors were proved to be independent risk factors of survival., Conclusion: RFA combined with TACE is an effective treatment for HCC with satisfactory short-term and long-term effects, especially for the patients with tumor of the size of 5.1 - 7 cm or multiple lesions.

Aim: To prepare specific antibodies against human PIWIL3 protein of Argonaute family and study its expression and distribution in human tumors., Methods: To synthesize the polypeptide using the optimal polypeptide immunogens based on sequence homology and peptide immunogenicty in the internal polypeptide selecting database. The synthesized polypeptide were combined with KLH, then immunized the experimental rabbits to produce antiserum. The antiserum was purified by gels custodited the polypeptide using affinity chromatograph, and validated its ability to combine with the corresponding proteins by enzyme-linked immunosorbentassay (ELISA) and Western blot. The expression and distribution of PIWIL3 protein were detected by human tumor tissue biochips., Results: The specific antibodies against human PIWIL3 proteins were prepared and had high purity. PIWIL3 proteins were expressed in the cytoplasma of human astrocytic glioma and meningioma., Conclusion: The specific antibodies against human PIWIL3 protein have been successfully prepared by using the internal polypeptide selecting database to obtain the optimal polypeptide immunogens, which are distinguished from other sequence homology proteins in the subfamily. These antibodies will have potential value for human PIWIL3 protein in human specific tumor pathogenesis.

Gene chip methods are applied to the study of gene expression. The differentially expressed genes in different specimens may be detected with parallel analysis by gene chip, which has greatly improved the traditional experiments in that only a single or several gene expressions need to be observed for each test, thereby speeding up the identification of differentially expressed genes and the construction of differential expression profiles. Many studies have applied this technology for Barrett's metaplasia and adenocarcinoma, and identified a number of candidate genes useful as biomarkers in cancer staging, prediction of recurrence, prognosis and treatment selection. This review described the gene expression profile and molecular changes related to Barrett's metaplasia and adenocarcinoma of the esophagus, with emphasis on its prognostic value and possibilities for targeted therapy in a clinical setting.

Objective: The expression of S100A6 (calcyclin), a member of the S100 calcium binding protein family, is elevated in a number of malignant tumors, but there have been few reports about its expression in gastric cancer. The aim of this study was to investigate its expression regulations in human gastric cancer and noncancerous mucosa, and the response to chemotherapeutic drugs in the gastric cancer cell line., Materials and Methods: In one matched gastric cancer sample pair, the serial analysis of gene expression (SAGE) experiment was conducted to compare the gene expression profiles between cancerous and adjacent tissues. To detect the expression regulations among more cancerous tissues, microarrays were carried out and real-time RT-PCR was conducted to validate the results. At the protein level, Western blot and tissue microarray (TMA) examination were further used to verify S100A6 expression. The regulation detection of S100A6 with flurouracil and doxorubicin at the mRNA and protein level was performed in the SGC7901 cell line., Results: With the SAGE strategy, five times more S100A6 tags were identified in cancer tissues than in normal tissues. With the cDNA microarray, S100A6 was found to be significantly upregulated in 21 of 42 (50%) nonselective gastric cancers. In 10 other paired samples, the upregulation of S100A6 was consolidated with RT-PCR and Western blot analysis as well. A total of 14 endoscopy-sectioned gastric noncancerous lesions and corresponding normal gastric mucosa were also applied to profile the gene expression; both cDNA microarray and RT-PCR demonstrated no significant alterations of S100A6 at the mRNA level. TMA examination showed that 34 of 52 (65.4%) cancer samples were positively stained, while only 17 of 80 (21.3%) noncancerous lesions were positively detected and all nine normal mucosae were detected to be negative. An in vitro experiment showed that in the gastric cell line SGC-7901, S100A6 mRNA was detected to be upregulated from 24 to 72 h after treatment with 5 mg/L 5-flurouracil or 0.3 mg/L doxorubicin, and there were two wave upregulations of the S100A6 protein., Conclusion: The observed regulated expression of S100A6 suggests that it is associated with gastric cancer tumorigenesis and quantitation of S100A6 is a promising tool for diagnosis of gastric cancer.

Objective: To find out the gene clusters associated with pulmonary embolism-deep venous thrombosis (PE-DVT) and elucidate the molecular genetic mechanism of PE., Methods: Peripheral venous blood samples were collected from 9 PE patients and 33 normal controls. The total RNA was extracted and purified. Nine PE cRNA probes labeled with cyanine 3 were constructed, and one standard cRNA probe labeled with cyanine 5 was synthesized based on the total RNA mixture of the controls. Hybridization with Agilent Whole Human Genome Oligo Microarray was performed. Eleven of the genes screened were randomly selected to be amplified by fluorescence quantitative PCR (FQ-PCR)., Results: 434 differential expression genes were screened from the 9 microarrays, including 36 gene transcripts. Associated with blood coagulation, 20 with immune or inflammatory response, 29 with metabolism, 26 with cell differentiation and apoptosis, 25 with cell growth/maintenance, 22 with cell-cell signaling or signal transduction, 14 with cytoskeleton or motility, 15 with ion channel or ion transport, 14 with transcription, and 6 with DNA/RNA binding. These 11 of these genes were randomly selected to be amplified by FQ-PCR. The differential expression of the 11 selected genes was consistent with the microarray., Conclusion: The differential expression of a lot of genes in the body may play a role in the process of initiation and development of PE-DVT.

Objective: To investigate NF-kappaB activity and the expression of phosphorylated p38 MAPK protein in lung tissue of acute paraquat poisoned rats and the effect of MT., Methods: One hundred and twenty-eight Sprague-Dawley (SD) rats were randomly divided into three experimental groups: poisoned group, MT group and control group. On the 1st, the 3rd, the 7th and the 14th day after exposure, levels of malondialdehyde (MDA) in serum were detected, NF-kappaB activity in the lung tissues was assessed by electrophoresis mobility shift assay (EMSA), the expression of the phosphorylated p38 MAPK was evaluated by Western blot method, the lung pathological changes of rats were observed., Results: The level of malondialdehyde (MDA) in serum increased significantly in poisoned group on the 1st day (4.45 +/- 1.23), the 3rd day (3.77 +/- 1.12) and the 7th day (2.84 +/- 0.96) nmol/ml compared with that in control group (1.36 +/- 0.52) nmol/ml (P < 0.01). There was a significant decrease in MT group on the 1st day (2.68 +/- 0.85), the 3rd day (1.97 +/- 0.74) and the 7th day (1.53 +/- 0.62) nmol/ml compared with poisoned group (P < 0.05). The expression of the phosphorylated p38 MAPK and NF-kappaB activity in lung tissue of poisoned group significantly increased compared with control group (P < 0.01). There was a significant decrease in NF-kappaB activity and expression of the phosphorylated p38 MAPK in the lung tissues in MT group compared with poisoned group (P < 0.05)., Conclusion: NF-kappaB and p38 MAPK could play an important role in lung injury of poisoned rats. MT may inhibit the expression of NF-kappaB and phosphorylated p38 MAPK, and therefore might have the therapeutical effect on acute paraquat poisoning.

Aim: To explore the molecular events taking place during human colon cancer development and progression through high-throughput tissue microarray analysis., Methods: We constructed two separate tissue microarrays containing 1.0 mm or 1.5 mm cylindrical samples acquired from 112 formalin-fixed and paraffin-embedded blocks, including carcinomas (n = 85), adenomatous polyps (n = 18), as well as normal para-cancerous colon tissues (n = 9). Immunohistochemical staining was applied to the analysis of the consecutive tissue microarray sections with antibodies for 11 different proteins, including p53, p21, bcl-2, bax, cyclin D1, PTEN, p-Akt1, beta-catenin, c-myc, nm23-h1 and Cox-2., Results: The protein expressions of p53, bcl-2, bax, cyclin D1, beta-catenin, c-myc, Cox-2 and nm23-h1 varied significantly among tissues from cancer, adenomatous polyps and normal colon mucosa (P = 0.003, P = 0.001, P = 0.000, P = 0.000, P = 0.034, P = 0.003, P = 0.002, and P = 0.007, respectively). Chi-square analysis showed that the statistically significant variables were p53, p21, bax, beta-catenin, c-myc, PTEN, p-Akt1, Cox-2 and nm23-h1 for histological grade (P = 0.005, P = 0.013, P = 0.044, P = 0.000, P = 0.000, P = 0.029, P = 0.000, P = 0.008, and P = 0.000, respectively), beta-catenin, c-myc and p-Akt1 for lymph node metastasis (P = 0.011, P = 0.005, and P = 0.032, respectively), beta-catenin, c-myc, Cox-2 and nm23-h1 for distance metastasis (P = 0.020, P = 0.000, P = 0.026, and P = 0.008, respectively), and cyclin D1, beta-catenin, c-myc, Cox-2 and nm23-h1 for clinical stages (P = 0.038, P = 0.008, P = 0.000, P = 0.016, and P = 0.014, respectively)., Conclusion: Tissue microarray immunohistochemical staining enables high-throughput analysis of genetic alterations contributing to human colon cancer development and progression. Our results implicate the potential roles of p53, cyclin D1, bcl-2, bax, Cox-2, beta-catenin and c-myc in development of human colon cancer and that of bcl-2, nm23-h1, PTEN and p-Akt1 in progression of human colon cancer.

Objectives: To analyze the differential expression genes (DEGs) among hepatocellular carcinoma (HCC), para-cancerous tissue (PCT) and normal liver tissue (NLT) and explore the target genes related to the development and progression of HCC., Methods: The total RNAs of matched HCC, PCT and NLT of HCC patients were isolated using one step Trizol method. Matched RNAs were qualified using 10 g/L agarose gel electrophoresis and lab-on-chip. cRNAs were synthesized, fluorescence labeled and purified after total RNAs were purified. The RNAs of HCC and NLT, HCC and PCT were hybridized with Agilent oligo microarray (21,074 probes). The fluorescence intensity features were detected by Agilent scanner and quantified by feature extraction software. The selected candidate genes were confirmed by SYBR Green I stained real time RT-PCR., Results: (1) The total RNA, reverse transcription product and fluorescence labeled cRNA were all of high quality; (2) There were 420 up-regulated genes and 552 down-regulated genes among 2-fold DEGs, including DKK1 (dickkopf homolog 1) which was 5-fold up-regulated; (3) The results of real time RT-PCR, using beta-actin as an internal control, showed that the 2-Delta Ct values of DKK1 in HCC, PCT and NLT were 0.089 504, 0.007,65 and 0.000,631 respectively., Conclusion: (1) The high throughput and effective Agilent oligo microarray can screen novel therapy targeted genes by analyzing the DEGs in development and progression of HCC; (2) The development and progression of HCC is a complicated process involving multigenes and multiprocedures; (3) DKK1, as a novel gene, is involved in the development and progression of HCC and may be a new therapy target.

Objective: To constructed tissue microarray containing specimens from gastric cancer and adjacent non-cancer tissue to survey the expression of p53, p16 and cyclooxygenase-2 (COX-2), and to obtain a comprehensive survey on the expression of three proteins in gastric cancer progression and their clinical significance., Methods: We constructed a tissue microarray containing 50 specimens from gastric cancer and 78 specimens from adjacent non-cancer tissue, and assayed three different proteins (p53, p16 and COX-2) by immunohistochemistry to consecutive formalin-fixed tissue microarray sections., Results: In non-cancer tissue, p53, p16 and COX-2 positive staining were 19%, 15% and 74% respectively. In gastric cancer tissue, p53, p16 and COX-2 positive staining were 50%, 54% and 94% respectively. There were a significant difference between two different tissues (P < 0.05). 52% (66/128) cases were COX-2+/p53-, only 1 case were COX-2-/p53+. There was a significant association between COX-2 and p53 (P < 0.05). COX-2 negative expression tissues were found significantly more often in p53-negative than in p53-positive cases. 52% (67/128) cases were COX-2+/p16-, only 1 case were COX-2-/p16+. A significant correlation was recognized between expression of COX-2 and p16 (P < 0.05). Logistic regression analysis revealed that the risk factors of tumorous histotype were p53, p16 and COX-2 positive expression together, determined by Wald chi2 test (P < 0.05; odds ratio, 18.889). There was reciprocal relationship among p53, p16 and COX-2., Conclusion: Combination analysis of p53, p16 and COX-2 may be useful for the prediction of gastric carcinogenesis.

Objective: To detect the expression of tumor suppressor protein p53, cyclin-dependent kinase inhibitor p16, and cyclooxygenase 2 (COX-2) in pancreatic cancer by tissue microarray and investigate the correlation among these three genes., Methods: 104 specimens of tissues, including pancreatic cancer tissue, non-cancer tissues not more than 1.5 cm from the cancer, and normal tissues, underwent microarray examination and immunohistochemistry to detect the expression of p53, p16, and COX-2. The correlation among these 3 genes was analyzed., Results: p53, p16, and COX-2 were all significantly highly expressed in the cancerous tissues in comparison with other tissues. P53 and p16 were both significantly correlated with COX-2 (both P < 0.05), however, there was not a correlation between p53 and p16 (P > 0.05). There was a reciprocal relationship between p53 and COX-2 (P < 0.05, OR = 19.686) influencing the pathogenesis of pancreatic cancer., Conclusion: Tissue microarray technique is effective method to detect multiple gene protein expression. Pathogenesis of pancreatic cancer is associated with p53, p16, and COX-2.

AIM:To investigate the expression of multiple genes and the behavior of cellular biology in gastric cancer (GC) and other gastric mucosal lesions and their relations to Helicobacter pylori (H. pylori) infection, tumor staging and histological subtypes.METHODS:Three hundred and twenty seven specimens of gastric mucosa obtained via endoscopy or surgical resection, and ABC immunohistochemical staining were used to detect the expression of p53, p16, Bcl-2 and COX-2 proteins.H. pylori was determined by rapid urea test combined with patholo-gical staining or 14 Curea breath test. Cellular image analysis was performed in 66 patients with intestinal metaplasia (IM) and/or dysplasia (Dys). In 30 of them, both cancer and the paracancerous tissues were obtained at the time of surgery. Histolo-gical pattern, tumor staging, lymph node metastasis, grading of differentiation and other clinical data were studied in the medical records.RESULTS:p16 expression of IM or Dys was significantly lower in positive H. pylori chronic atrophic gastritis (CAG) than those with negative H. pylori (CAG: 54.8% vs 88.0%, IM:34.4% vs 69.6%, Dys: 23.8% vs 53.6%, all P < 0.05), Bcl-2 or COX-2 expression of IM or Dys in positive H. pylori cases was signi-ficantly higher than that without H. pylori (Bcl-2: 68.8% vs 23.9%, 90.5% vs 60.7%; COX-2: 50.0% vs 10.8%, 61.8% vs 17.8%; all P <0.05). The mean number of most parame-ters of cellular image analysis in positive H. pylori group was significantly higher than that in negative H. pylori group (Ellipser: 53 plus minus 14, 40 plus minus 12&mgr;m, Area(1): 748 plus minus 572, 302 plus minus 202&mgr;m(2), Area(2): 3050 plus minus 1661, 1681 plus minus 1990&mgr;m(2), all P< 0.05; Ellipseb: 79 plus minus 23, 58 plus minus 15&mgr;m, Ratio-1: 22% plus minus5%,13% plus minus4%,Ratio-2:79% plus minus17%,53% plus minus20%,all P<0.01). There was significant correl-ation between Bcl-2 and histologic pattern of gastric carcinoma, and between COX-2 and tumor staging or lymph node metasta sis (Bcl-2: 75.0% vs16.7%; COX-2: 76.0% vs 20.0%, 79.2% vs 16.7%; all P< 0.05).CONCLUSION:p16, Bcl-2, and COX-2 but not p53 gene may play a role in the early genesis/progression of gastric carcinoma and are associated with H. pylori infection. p53 gene is relatively late event in gastric tumorigenesis and mainly relates to its progression. There is more cellular-biological behavior of malignant tumor in gastric mucosal lesions with H. pylori infec-tion. Aberrant Bcl-2 protein expression appears to be preferentially associated with the intestinal type cancer. COX-2 seems to be related to tumor staging and lymph node metastasis.