Objective To investigate the influence of long non-coding RNA HOTAIR in proliferation and apoptosis of human tongue squamous cell carcinoma in vitro and in vivo. Methods siHOTAIR was used to inhibit the HOTAIR expression in Tb3.1 human tongue squamous cell carcinoma cell line. The experiments were divided into siHOTAIR group, nonsense sequence group and blank control group. Real-time PCR was used to detect the HOTAIR expression. MTT assay was employed to determine the cell survival. The expression levels of Bcl2, BAX, caspase-3, cleaved caspase-3 were examined by Western blot assay. Tb3.1 xenograft tumor model was established in BALB/c nude mice, and the tumor model was divided into control group, negative group, and siHOTAIR treated group. The tumor tissues were measured by immunohistochemistry stain (IHC) and TUNEL assay. Results The detection of real-time PCR showed that HOTAIR expression was reduced after treated with siHOTAIR. Western blots assay showed that Bcl-2 protein was suppressed while cleaved caspase-3 and BAX protein were up-regulated after treated with siHOTAIR. MTT assay indicated that the cell survival rate was significantly reduced in siHOTAIR treated group. Flow cytometry detected that apoptosis levels were increased in siHOTAIR group. The level of cell senescence was higher in the siHOTAIR group than that of control group. Results of IHC indicated that Ki-67 and Bcl-2 protein of tumor tissue were inhibited, while BAX and cleaved caspase-3protein expressions were elevated simultaneously in the siHOTAIR group. TUNEL assay suggested that more apoptosis was observed in siHOTAIR group. Conclusion HOTAIR can affect proliferation and apoptosis of tongue squamous cancer cells. HOTAIR may be one of the new candidate targets for human tongue cancer therapy.

Objective To evaluate the factors influencing early diagnosis and prognosis in patients with pancreatic carcinoma.Methods The clinical data of 280 patients who had complete follow-up data with pancreatic carcinoma treated from January 2002 to January 2007 were reviewed retrospectively.The medical history and follow-up data were collected from all patients.Survival rate was calculated by the life table method and the Kaplan-Meier estimation.Log-rank test was used for univariate prognostic analysis and Cox regression was used for multivariate prognostic analysis.Results 91.8%of the patients were more than 40 years old and the peak age was 50～73 years old;the major presentations were abdominal pain and jaundice.Major imaging tests included B-ultrasound and CT,the sensitivity was 70.6%,95.3%,respectively;89.3%of patients had combined B-ultrasound and CT examination.The sensitivity of CA19-9 was 81.1%.The median survival time was(7.0±0.5)months.Overall survival rates at 1～5 year survival rates were 28%,9%,6%,2%,and 1%.Univariate analysis suggested that age＞65 years old,CA19-9＞mean value,TNM Ⅲ or Ⅳ stage,lymph nodes invasion,vascular invasion,and metastasis of two or more organs,non-surgical treatment,KPS score＜60 points,weight loss≥5 kg were poor prognostic factors;Cox multivariate analysis showed that treatment modalities,age,TNM stage,KPS score and ascites were independent risk factors for dismal prognosis.Conclusions The age,ascites,tumor stage and treatment modalities affected the prognosis of patients with pancreatic cancer.Early diagnosis and treatment was important to improve the survival time of patients with pancreatic cancer.

To investigate the effects of manual acupuncture at Shenmen (HT7) or Taiyuan (LU9) on the attention function of the brain, and to lay an experimental foundation for researching brain function and integration mechanisms of the human brain in relation to acupuncture stimulation.

Objective To explore the expressions of Cyclin-dependent kinase 5 (CDK5) and Epithelial-Mesenchymal Transition (EMT) related proteins including N-cadherin, Vimentin and E-cadherin in head and neck squamous cell carcino? ma (HNSCC), and to determine the relationship between the expression of CDK5 and prognosis. Methods The expression levels of CDK5 and EMT related proteins were evaluated by immunohistochemistry in 55 patients who were diagnosed as HN?SCC. They were also analyzed in different clinical pathological factors. The correlation of CDK5 and EMT related proteins as well as the relationship between the expression of CDK5 and prognosis were also analyzed. Results The expression level of CDK5 was significantly higher in patients with lymph node metastasis than that in patients with non-lymph node metastasis (91.67%vs 30.23%, P<0.05). It’s also higher in T3-T4 stages than that in T1-T2 stages (85%vs 20%, P<0.05). The ex?pression levels of N-cadherin and Vimentin were significantly higher in patients with lymph node metastasis than those in patients with non-lymph node metastasis (75.00%vs 6.98%;91.67%vs 27.91%, all P<0.05). However, the expression level of E-cadherin was significantly lower in patients with lymph node metastasis (8.33%vs 86.05%, P<0.05) compared to that in patients without. CDK5 was positively correlated with N-cadherin and Vimentin, but negatively correlated with E-cad?herin (rs=0.512, 0.443,-0.363, all P<0.01). The 3-year survival rates were significantly lower in patients with high expres?sion of CDK5 (37.5%) than that in patients with low expression of CDK5 (87%, Log-rankχ2=12.678, P<0.01). Conclusion CDK5 and EMT related proteins were activated abnormally in HNSCC with lymph node metastasis. CDK5 may be a new bio?logical marker for prognosis of HNSCC.

Objective To study the influence of the small interfering RNA (siRNA) interference TMPRSS4 expression on human pancreatic cancer SW1990 cell's proliferation and invasion. Methods The four eukaryotic expression vector of TMPRSS4 gene were synthesized in vitro and were transfected transiently into human pancreatic cancer SW1990 cells. TMPRSS4 mRNA expression of transfected cells was detected by real-time RT-PCR. The most efficient eukaryotic expression vector was used to be transfected into SW1990 cells. By using G418, cell strain that can silence TMPRSS4 gene stably was screened. The TMPRSS4 mRNA expression of the stable cell strain was detected by real time PCR TMPRSS4 protein expression was detected by western blot. The proliferation ability of transfected SW1990 cells was detected by CCK-8 method. By Transwell, the invasion change of SW1990 cell was detected. Results A stable cell strain, SW1990/psi TMPRSS4, was successfully constructed, in which the expression level of TMPRSS4 could be reduced stably by RNA interference. Cell transfection efficiency was 82.9%. Compared with the control group, the TMPRSS4 mRNA and protein levels were reduced by 80.1% and 60% ,and number of penetrating cells was 118.6 ±13.4 in SW1990/psi TMPRSS4 group, which was significantly lower than those in the negative control group (157.4 ± 12.9) and control group (157.0±9.5, P <0.01). Cells invasion inhibitory rate was 24.5% in SW1990/psi TMPRSS4 group. The cell proliferation was not significantly different among all the groups. Conclusions A stable cell strain is screened successfully in which the expression level of TMPRSS4 can be reduced stably. The down-regulation of TMPRSS4 gene expression level can inhibit the invasion of SW1990 cells, but has no effect on cell proliferation.

OBJECTIVETo assess the value of routine intra-aortic balloon pump (IABP) support in patients with high-risk acute myocardial infarction (AMI) undergoing percutaneous coronary intervention (PCI).

METHODSThe clinical data of 41 patients with high-risk AMI undergoing emergency PCI with routine IABP support were retrospectively reviewed, and 38 patients paired with the former group receiving emergency PCI for high-risk AMI without IABP support at the same time were included as the control group. Thirty days after the operation, the two groups were compared for myocardial ischemic events, left ventricular function and major adverse cardiac events (MACE).

RESULTSPatients receiving IABP support had a significantly lower incidence of myocardial ischemic events than those without IABP (4.9% vs 15.8%, P<0.05), and showed greater improvement in the left ventricular function. Significant differences were also observed in the mortality rate, incidence of reinfarction and revascularization rate between the two groups, but not in the rate of MACE.

CONCLUSIONPatients undergoing PCI for high-risk acute AMI can benefit from routine IABP support in terms of improvement of left ventricular function and reduce myocardial ischemic events and the rate of MACE. These results, however, still await further confirmation by large-scale clinical trials.

Angioplasty, Balloon, Coronary ; Female ; Humans ; Incidence ; Intra-Aortic Balloon Pumping ; methods ; Male ; Middle Aged ; Myocardial Infarction ; mortality ; therapy ; Myocardial Ischemia ; Retrospective Studies ; Treatment Outcome ; Ventricular Function, Left

OBJECTIVETo evaluate the effects of portal vein microscopic and macroscopic tumor thrombi on post-operation patients with hepatocellular carcinoma (HCC).

METHODSThree thousand three hundred and forty eight HCC patients were retrospectively reviewed, which were divided into no portal vein tumor thrombi (PVTT), microscopic PVTT and macroscopic PVTT groups according to the pathology, effects of portal vein microscopic and macroscopic tumor thrombi on post-operation patients's survival were studied by univariate analysis and overall survival was evaluated in each group.

RESULTSHazard ratio (HR) of portal vein microscopic tumor thrombi and macroscopic tumor thrombi was 1.421 and 3.136 respectively; The overall 1-, 3-, 5- and 10-year cumulative survival rate was 85.97%, 62.78%, 49.88% and 35.42% respectively, and mean time for survival was 59.7 months in group without PVTT, while 74.42%, 51.66%, 39.25% and 27.28% respectively and mean time for survival 39.1 months in group with microscopic PVTT, 52.59%, 25.97%, 20.42% and 11.33% respectively and mean time for survival 13.5 months in group with macroscopic PVTT.

CONCLUSIONSPVTT was an important prognostic factor for survival in post-operation patients with HCC while macroscopic PVTT was more danger than microscopic PVTT. The period of microscopic PVTT was the landmark affecting post-operation survival.

Carcinoma, Hepatocellular ; mortality ; pathology ; surgery ; Humans ; Liver Neoplasms ; mortality ; pathology ; surgery ; Neoplastic Cells, Circulating ; Portal Vein ; pathology ; Retrospective Studies ; Survival Rate

OBJECTIVETo identify the metastasis-related miRNAs in hepatocellular carcinoma (HCC) cell lines.

METHODSA qRT-PCR method was established and optimized.

RESULTSAll candidate metastasis associated miRNAs except miR-124a were expressed in high metastasis cell line MHCC97H and low metastasis cell line MHCC97L, while some miRNAs were differentially expressed between liver cancer cell line (HepG2) and hepatic cell line (L02) (P less than 0.05), these miRNAs include: miR-148b (1.96+/-0.51 vs 3.76+/-0.28), miR-9 (-4.38+/-0.86 vs -1.10+/-0.53), miR-30c (8.41+/-0.40 vs 6.82+/-0.29), miR-338 (3.14+/-0.29 vs -2.36+/-0.32), miR-34a (0.71+/-0.40 vs -2.95+/-0.26), Let-7g (-4.07+/-0.55 vs -6.98+/-0.56). miR-148b expression was about 4 times higher than miR-148a [5.46 (IQR 4.25-6.67) vs 1.29 (IQR 0.94-1.64)] in all cell line tested (Z=-5.097, P=3x10(-7)).

CONCLUSIONThis study may help to understand the biological significance of miRNAs in HCC metastasis.

Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line ; Cell Line, Tumor ; DNA, Complementary ; genetics ; Epithelial Cells ; metabolism ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; MicroRNAs ; genetics ; metabolism ; Neoplasm Metastasis ; Polymerase Chain Reaction

OBJECTIVETo screen hepatocellular carcinoma (HCC) autoantibodies as diagnostic biomarkers or therapy targets by serologic proteome analysis (SERPA).

METHODSTotal proteins extracted from human HCC cell line HCCLM3 were separated by two-dimensional electrophoresis (2-DE) and then transferred onto PVDF membranes, which were subsequently incubated with sera from HCC, hepatitis B virus (HBV) infected patients or healthy volunteers. All immuno-reactive protein spots on blot films were matched to those on 2-DE gel maps by image analysis and identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS/MS).

RESULTS2-DE gel maps of HCCLM3 and corresponding blot films of good quality and reproducibility were established. The number of spots on HCCLM3 2-DE reference gel totaled 603 and those on HCC, HBV and healthy sera blotted films were 70.75+/-24.25, 68.5+/-23.44 and 41.38+/-15.05, respectively. Blot films of HCC and HBV groups had more spots than those of the healthy group (P < 0.05) while no significance was found between films of HCC and HBV groups. By identification, those HCC autoantibodies could be classified as nuclear proteins, cytoskeleton proteins, heat shock proteins and metabolic enzymes.

CONCLUSIONSerological proteome analysis is a high throughput technique for screening tumor autoantibodies. Those newly identified HCC associated tumor antigens and corresponding autoantibodies can be used in the early diagnosis or immuno-therapy of HCC.

Antibodies, Neoplasm ; analysis ; Autoantibodies ; analysis ; Carcinoma, Hepatocellular ; immunology ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Liver Neoplasms ; immunology ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Tumor Cells, Cultured

OBJECTIVETo clarify three-grade criteria of curative resection for primary liver cancer (PLC) and evaluate their clinical significance.

METHODSCriteria of curative resection of PLC were summed up to three grades. Grade I: complete removal of all gross tumors with no residual tumor at the excision margin. Grade II: on the basis of Grade I, there was no extrahepatic metastasis, no hilar lymph node metastasis, no tumor thrombus in the main trunks and their primary tributaries of the portal vein, common hepatic duct, hepatic vein and vena cava inferior, and the tumor was not more than two in number. Grade III: in addition to the above criteria, AFP dropped to normal level (in patients with elevated AFP before surgery) within 2 months after operation, and no residual tumor upon diagnostic imaging. A total of 354 cases with PLC who had their liver resected was reviewed. Patients in each grade were divided into two portions depending on whether the treatment was curative or palliative.

RESULTSThe survival of patients receiving curative treatment was better than those receiving palliative treatment (P < 0.01). This was true for patients whose treatment belonged to anyone of the three-grade criteria. The survival was improved along with the promotion of curative criteria used. The 5-year survival rate of Grade I, II and III patients undergone curative resection was 43.2%, 51.2% and 64.4%, respectively (P < 0.01).

CONCLUSION1. The three-grade criteria may be used for judging the radicality of tumor resection for PLC. 2. The more stringent the criteria used, the better the survival would be. 3. Adopting high-grade criteria to select cases, to guide operation and postoperative follow-up would improve the results of liver resection for PLC.

Female ; Hepatectomy ; methods ; Humans ; Liver Neoplasms ; mortality ; surgery ; Male ; Middle Aged ; Survival Rate