Objective:To investigate the correlation of FAT10 expression with the malignant characteristics of hepatocellular car-cinoma (HCC), and to explore the effect of FAT10 on RhoA and cytoskeleton of HCC. Methods: Immunohistochemistry (IHC) was used to detect the FAT10 expression level of 108 HCC patients, and the correlation between the expression of FAT10 and the malignant characteristics of HCC patients was analyzed. We transiently transfected plasmids with overexpressed FAT10 using 7721 and HepG2 cells or interfered with FAT10 expression using siRNA in Huh7 and LM3 cells. Active-RhoA, total-RhoA, and ROCK protein expres-sion levels were detected by Western blot analysis after overexpression or interference. We also used immunofluorescence to detect changes in the cytoskeleton protein F-actin after FAT10 overexpression in 7721 cells. Results:Correlation analysis showed that both ac-tive-RhoA and FAT10 expression levels were significantly correlated with clinical malignant characteristics by using IHC (RhoA:me-tastasis, P=0.036 and recurrence, P=0.026;FAT10:metastasis, P=0.031 and recurrence P=0.004). In addition, active-RhoA expression level was correlated with FAT10 (P=0.000). Survival analysis showed that the prognoses of low-expression active RhoA (P=0.019) or FAT10 (P=0.026) groups were significantly better than those of the high-expression groups. Western blot analysis showed that FAT10 increased the expression of active-RhoA and ROCK. However, the expression of active-RhoA and ROCK decreased after FAT10 inter-ference. F-actin expression increased in the 7721 cells with overexpressed FAT10 (all P<0.01). Moreover, FAT10 facilitated F-actin ag-gregation on cell membrane and changes in F-actin. Conclusion:FAT10 is correlated with the malignant characteristics of HCC and may promote changes in HCC cytoskeleton induced by active-RhoA.

Objective: To evaluate the safety and efficacy of nonmyeloablative preconditioning allogeneic hematopoietic stem cell transplantation (NMHSCT) in the therapy of unidentified relapse and primary solid tu-mors, and to study the anti-tumor immunity induced by the effect of Graft-Verus-Tumor (GVT). Methods: A to-tal of 13 difficult-to-treat cancer patients received NMSCT and the efficacy and side effects were observed. Re-sults: One case had CR, 2 cases had PR, 4 cases had SD, 5 cases had PD, and 1 case died of complica-tions associated with transplantation. One case was GVT (+++), 3 cases were GVT (++), 5 cases were GVT(+), and 4 cases were GVT (-). The main side effect was acute GVHD presented as diarrhea and infec-tion. VOD and brain disease were rare. Conclusion: Nonmyeloablative allogeneic stem cell transplantation is safe and effective for solide tumors.

Objective:To transfect the non-viral vector polyethylenimine (PEI)mediated miR-2861 mimic into the MC3T3-E1 cell line,and to explore the transfection efficiency of PEI/miR-2861 complex and its effects on the proliferation and osteogenesis differentiation in pre-osteoblasts. Methods:The proper amount of PEI was blended with miR-2861 mimic and negative control (NC)separately in a ratio of N∶P=10∶1,and they were divided into experiment group and NC group. The NC/PEI complex acted as the NC group was used to eliminate the interference of osteogenesis from the addition of double-stranded RNA mimic.MTT assay was used to determine the optimal concentration of PEI/miR-2861 mimic complex.The fluorescence imaging technique and bulge-loop RT-PCR were used to detect the transfection efficiency and mRNA expression of miRNA-2861 in the cells with different concentrations (10,30, 50,and 100 nmol · L-1 ), separately.The osteogenesis ability of MC3T3 cells was identified with RT-PCR and Alizarin red staining with the selected concentration of PEI/miR-2861 by transient transfection.Results:Compared with blank control group,the proliferation rates of MC3T3 cells in 100 nmol·L-1 PEI/miR-2861 group was decreased significantly at 72 h (P < 0. 05 ). With the increasing of transfected concentration the transfection efficiency of miRNA/PEI complex was increased gradually.The results of Alizarin red staining and quantitative analysis showed that calcium deposits were more and bigger in experiment group after induced for 21 d,while both in blank control group and NC group they were less.Conclusion:The miRNA-2861 mimic can be effectively transfected into the MC3T3-E1 cell line and expresses with a high level,which is mediated by PEI as the gene vector.miR-2861 mimic has a certain ability of promoting osteogenesis differentiation of MC3T3-E1 cells.

OBJECTIVETo investigate the expression of casein kinase 2β in esophageal carcinoma tissues and analyze its clinical significance.

METHODSThe expression of CK2β in a tissue chip containing 60 normal esophageal mucosa and 60 colorectal cancer specimens were detected immunohistochemically. Ten human esophageal carcinoma and adjacent normal tissues were examined for the expression of CK2β protein and mRNA using Western blotting and real-time quantitative PCR, respectively.

RESULTSA strong expression of CK2β was found in 85.71% of the esophageal cancer tissues; 1.79% of the cancer tissues were negative for CK2β expression, and 1.79% and 10.71% of the cancer tissues were weakly and moderately positive, respectively. In the normal mucosal tissues, 96.67% of the tissues were negative for CK2β and 3.33% showed weak CK2β expression, showing a significant difference in the distribution of CK2β between normal and esophageal carcinoma tissues (P<0.001). The expression level of CK2β in esophageal cancers was associated with the pathological stage (TNM) (P=0.010). Western blotting and real-time quantitative PCR also confirmed an increased CK2β expression in the esophageal cancer tissues.

CONCLUSIONThe high expression of protein kinase CK2β is closely related to the carcinogenesis and malignancy of esophageal cancer.

Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Casein Kinase II ; metabolism ; Esophageal Neoplasms ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Middle Aged ; Neoplasm Staging

OBJECTIVE To investigate the surgical methods for the lesions involved the common carotid artery.METHODS The clinical data of 11 cases with lesions involved the common carotid artery who underwent operations were retrospectively studied.The lesions were 1 case with recurrence tumor after 3/4 partial laryngectomy,1 case with bleeding of the carotid aneurysm caused by tuberculosis,1 case with iatrogenic carotid aneurysm,3 cases with carotid body tumor,1 case with thyroid gland cancer,2 cases with neck tumor,1 case with injury of the carotid artery and 1 case with gas gangrene.RESULTS Common carotid artery was reconstructed in 2 cases after removal of the tumors.The tumors were resected using the carotid shunt in 2 cases.Common carotid artery was sutured in 1 case with neck injury.The common carotid artery was repaired in 1 case with iatrogenic carotid aneurysm after removal of the tumor.The carotid artery was dissected out from the thyroid gland cancer in 1 case.The common carotid artery was reserved in 2 cases after resection of the neck tumors. Neck drainage was performed in the case with gas gangrene.CONCLUSION The surgical methods for lesions involved the carotid artery after removal of the tumors include the reconstruction of the carotid artery, resection and suture the carotid artery,and free of the carotid artery from the tumors.

Cell culture is an important tool for biological research.To better understand the pathogenesis and therapeutic methods of the tumors,the three-dimensional cell culture is applied by more and more researchers to create a culture environment that closes to the tumor microoenvironment.Thanks to the advances in the tissue engineering technology,many kinds of models of the three-dimensional cell culture achieve wide accessibility.Compared with the traditional two-dimensional cell culture,the three-dimensional cell culture is better in simulating physiological features of the human histology and cells,including cell proliferation and differentiation,the interaction of cell to cell and cell to matrix.This paper reviews the progress of multi-cellular tumor spheroids (MCTS) culture technique of the three-dimensional cell culture for treatment of bladder cancer.

OBJECTIVE: To explore the role of Long noncoding RNA UFC1 (lincRNA-UFC1) in modulating the metastasis and invasion of hepatocellular carcinoma (HCC) cells and the underlying mechanism. METHODS: Human HCC cell line Huh7 was infected with the lentiviral vector carrying lincRNA-UFC1 to obtain a cell line with lincRNA-UFC1 overexpression. A short hairpin RNA (shRNA) targeting lincRNA-UFC1 was delivered in human HCC BEL-7402 cells via a lentiviral vector to obtain a cell line with lincRNA-UFC1 knockdown. Expression levels of lincRNA-UFC1 in the two HCC cell lines were detected using real-time PCR, and the changes in the cell invasion and migration in response to lincRNA-UFC1 overexpression or knockdown were analyzed using Transwell and wound-healing assays. The expressions of GSK-3β/β-catenin-related proteins in the cells were detected with Western blotting. XAV-939, a GSK-3β/β-catenin inhibitor, was used for assessing the impact of lincRNAUFC1 overexpression on the invasion and migration of the HCC cells through Transwell and wound-healing assays. RESULTS: Overexpression of lincRNA-UFC1 significantly promoted the invasion and migration of Huh7 cells as compared with the control cells ( < 0.001), while lincRNA-UFC1 knockdown obviously suppressed the invasion and migration of BEL-7402 cells ( < 0.001). The results of Western blotting showed that the expressions of proteins associated with the cell invasion and migration, namely β-catenin and P-GSK-3β, were significantly upregulated in response to lincRNA-UFC1 overexpression, and were obviously lowered after lincRNA-UFC1 knockdown. Treatment of the cells with XAV-939 significantly reversed the effect of lincRNA-UFC1 overexpression on the cell invasion and migration ( < 0.001). CONCLUSIONS lincRNA-UFC1 overexpresison promotes cell invasion and migration through the GSK-3β/β-catenin axis in HCC cells .
Carcinoma, Hepatocellular ; genetics ; Cell Line, Tumor ; Glycogen Synthase Kinase 3 beta ; Humans ; Liver Neoplasms ; genetics ; Neoplasm Invasiveness ; Neoplasm Metastasis ; RNA, Long Noncoding ; beta Catenin

A new method for detection of Escherichia coli exist in licorice decoction was developed by using DNA-based electrochemical biosensor. The thiolated capture probe was immobilized on a gold electrode at first. Then the aptamer for Escherichia coli was combined with the capture probe by hybridization. Due to the stronger interaction between the aptamer and the E. coli, the aptamer can dissociate from the capture probe in the presence of E. coli in licorice decoction. The biotinylated detection probe was hybridized with the single-strand capture probe. As a result, the electrochemical response to Escherichia coli can be measured by using differential pulse voltammetric in the presence of α-naphthyl phosphate. The plot of peak current vs. the logarithm of concentration in the range from 2.7×10² to 2.7×10⁸ CFU·mL⁻¹ displayed a linear relationship with a detection limit of 50 CFU·mL⁻¹. The relative standard deviation of 3 successive scans was 2.5%，2.1%，4.6% for 2×10²，2×10⁴，2×106：⁶ CFU·mL⁻¹ E. coli, respectively. The proposed procedure showed better specificity to E. coli in comparison to Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus subtilis. In the detection of the real extractum glycyrrhizae, the results between the proposed strategy and the GB assay showed high degree of agreement, demonstrating the designed biosensor could be utilized as a powerful tool for microbial examination for traditional Chinese medicine.

OBJECTIVE: To screen the key genes related to the prognosis of lung adenocarcinoma through big data analysis and explore their clinical value and potential mechanism. METHODS: We analyzed GSE18842, GSE27262, and GSE33532 gene expression profile data obtained from the Gene Expression Omnibus (GEO). Bioinformatics methods were used to screen the differentially expressed genes in lung adenocarcinoma tissues and KEGG and GO enrichment analysis was performed, followed by PPI interaction network analysis, module analysis, differential expression analysis, and prognosis analysis. The expressions of MAD2L1 and TTK by immunohistochemistry were verified in 35 non-small cell lung cancer specimens and paired adjacent tissues. RESULTS: We identified a total of 256 genes that showed significant differential expressions in lung adenocarcinoma, including 66 up-regulated and 190 down-regulated genes. Thirty-two up-regulated core genes were screened by functional analysis, and among them 29 were shown to significantly correlate with a poor prognosis of patients with lung adenocarcinoma. All the 29 genes were highly expressed in lung adenocarcinoma tissues compared with normal lung tissues and were mainly enriched in cell cycle pathways. Seven of these key genes were closely related to the spindle assembly checkpoint (SAC) complex and responsible for regulating cell behavior in G2/M phase. We selected SAC-related proteins TTK and MAD2L1 to test their expressions in clinical tumor samples, and detected their overexpression in lung adenocarcinoma tissues as compared with the adjacent tissues. CONCLUSIONS Seven SAC complex-related genes, including TTK and MAD2L1, are overexpressed in lung adenocarcinoma tissues with close correlation with the prognosis of the patients.
Adenocarcinoma of Lung/genetics* ; Big Data ; Cell Cycle Proteins/genetics* ; Computational Biology ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms/genetics* ; M Phase Cell Cycle Checkpoints ; Mad2 Proteins/genetics* ; Protein-Serine-Threonine Kinases/genetics* ; Protein-Tyrosine Kinases/genetics*

OBJECTIVETo identify clinical characteristics of hospitalized laboratory-confirmed influenza cases of children under 15 years old, and their risk factors of influenza infection.

METHODSAnalyzing the reports of hospitalized laboratory-confirmed influenza cases of children under 15 years old who were detected by the sentinel surveillance systems in 10 provinces from December 2009 to June 2014. Such data as their demographic, medical history, clinical symptoms and signs, treatment and outcome were collected using questionnaires, with their clinical characteristics and their risk factors of influenza infection described.

RESULTSOf the 2 937 severe acute respiratory infection inpatients, 190 (6.5%) were laboratory-confirmed influenza cases. 123 (64.7%) of such confirmed cases were male, and 139 (73.2%) were children under 5 years old, with age median of 3.0 years (IQR: 1.0-5.0 years). 20 (10.5%) of them had at least one chronic medical condition, mostly chronic cardiovascular disease (3.2%), immunosuppressive disease (3.2%), and cancer/tumor (2.6%). Most common clinical symptoms of the cases were fever (92.6%) and cough (88.8%), of which abnormal pulmonary auscultation (51.1%) and abnormal chest X-ray performance (36.1%) were the most common clinical signs. 29 cases (15.8%) had complications, of which pneumonia (15.3%) was most common. 16 cases (8.6%) used antiviral drugs, and 4 cases (2.2%) were admitted into ICU. Risk factor analysis suggested that age < 6 months (OR = 0.406, 95% CI: 0.203-0.815) was a protective factor against influenza infection; and age 5-9 years old (OR = 2.535, 95% CI: 1.059-6.066) was a risk factor for influenza infection.

CONCLUSIONHospitalized laboratory-confirmed influenza cases were found mostly in children under 5 years old. Risk exposure for influenza infection varied among age groups.

Acute Disease ; Adolescent ; Antiviral Agents ; Child ; Child, Preschool ; China ; epidemiology ; Cough ; Female ; Fever ; Hospitalization ; Humans ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; epidemiology ; pathology ; Inpatients ; Laboratories ; Male ; Protective Factors ; Risk Assessment ; Risk Factors ; Sentinel Surveillance ; Surveys and Questionnaires