10.3969/j.issn.2095-4344.2013.26.022

Objective To investigate the inhibitory effect of siRNA human telomerase transcriptase (hTERT) on activity of telomerase and proliferation of lung carcinoma cell A549. Methods A plasmid including U6 promoter and siRNA of hTERT was designed and constructed. The plasmid was transfected into A549 cell line. The telomerase activity was tested by telomerase repeat amplification protocol ELISA (TRAP-ELISA). MTT assay was used to assay the cell proliferation activity,and hTERT expression was assessed by Western blot. Result The U6 expression plasmid that was constructed for hTERT gene 745 showed obvious interfering effect. hTERT-siRNA could down-regulate the expression of hTERT protein,inhibit telomerase activity and proliferation of A549 cells. Conclusion siRNA of hTERT can inhibit the expression of human telomerase and proliferation of A549 cells. It may open a new approach to the use of siRNA as a new tool to study gene function in cancer cell lines,and may be developed to be a new gene therapeutic agent for cancer.

Hepatocellular carcinoma is a major disease for serious threat to human health, ranked third in the world of cancer causes of death and second in China' s in cancer mortality. There are gender differences in the incidence of HCC and numerous studies on estrogen' s role in the formation of HCC, and the formation of estrogen in the liver plays a protective role during the hepatocarcinogenesis. Some studies have shown estrogen inhibiting the expression of interleukin-6 thereby reducing the incidence of HCC. IL-6 and HGF have an important role in promoting HCC formation process, estrogen and these factors have are also important links. Now require in-depth exploration of novel adjuvant therapy is required to improve the survival rate of patients with HCC.

Objective To study the efficacy and safety of reboxetine and citalopram in treating retardant depression.Methods A total of 59 patients with retardant depression were randomly divided into reboxetine group(n=29)and citalopram group(n=30)for the treatment of 6 weeks.The patients were assessed with the Hamilton depression scale(HAMD)and treatment emergent symptom scale(TESS).Results At the end of 6 weeks,the total scores of HAMD in reboxetine(18.48±6.38)group and citalopram(11.60±4.46)group both significantly decreased(P＜0.001).Between two groups,the efficacy(82.8% and 56.7%)had significant differences(P＜0.05)but side effects had no statistical significance.Conclusion Reboxetine has significant curative efficacy,milder adverse effects and higher safety for treating patients with retardant depression.

Objective To analyze the gene expression profiles in response to ΔNp63α overexpression, and screen the potential target genes or signal pathways regulated by ΔNp63α. Methods To generate ΔNp63α overexpressed SiHa cells ( SiHa-ΔNp63α) and the control cells ( SiHa-NC) , recombinant lentivirus transfection was performed. Microarray was applied to detect the change of gene expression profiles, and the results were analyzed with bioinfor-matic software. Quantitative real-time PCR was used to validate the expression levels of selected genes. Results Among the 1405 differentially expressed genes which were statistically significant, >1. 5 fold increase or reduce of gene expression, 843 were up-regulated and 562 were down-regulated in SiHa cells with ΔNp63α overexpression. The genes were mostly involved in cell development,cycle regulation, signal transduction, communication, adhe-sion, metastasis and invasion, etc. The involved signal pathways consisted of antigen processing and presentation, cytokine-cytokine receptor interaction, cell adhesion, complement and coagulation cascades, and so on. Conclu-sion The research on the potential target genes or mediated signal pathways regulated by ΔNp63α could be helpful to explain the development of cervical cancer.

Objective To investigate the effects of anti-sense hTERT on the expression of VEGF and the receptor in SGC7901 cells, and study the influence in angiogenesis and progression in gastric carcinoma. Methods SGC7901 cell line was transfected by the recombinant virus containing sense and anti-sense hTERT cDNA. Then the expression of VEGF-C and Flt-4 was observed with RT-PCR, distribution of cell cycles was determined with flow cytometry. The expression of VEGF-C and Flt-4 protein was assessed with immunohistochemistry. Result The distribution of cell cycle of antisense hTERT transfected SGC7901 cells was changed, and the mRNA and protein expression of VEGF-C and Flt-4 was significantly down-regulated. Conclusion Anti-sense hTERT can act as an agent for inhibiting VEGF-C and Flt-4 mRNA and protein level, and it blocks tumor angiogenesis and lymphogenous metastasis.

The CD19-targeted chimeric antigen receptor (CAR) T-cell treatment has achieved clinical success in treating B-cell lineage hematopoietic malignancies. This accomplishment involved the precise and efficient elimination of tumor cells by tumor-associated an-tigen-redirected immune cells. As a result, the reinitiation of several CAR T-cell (CART) based clinical trials in solid tumors was promot-ed. However, developing a feasible and efficient CAR T based therapeutic modality for solid tumors is urgently needed given the differ-ential properties between liquid and solid tumors. In this review, we discuss the advances in the management of cytotherapeutic mo-dality for solid tumors. The aspects considered include toxicity management, target selection, and primer or conditioning treatment.

Objective　To investigate the efficacy of combined use of apigenin and resveratrol in the treatment of non-alcoholic fatty liver disease (NAFLD). Methods　Fifty male C57BL/6 mice were randomly divided into a normal group (n=10) and a model group (n=40). The NAFLD model was established in the model group using carbon tetrachloride (CCl4) and a high-fat diet. After successful modeling, the model group was further divided into a model group, an apigenin group, a resveratrol group, and an apigenin and resveratrol combined group (combined administration group), with 10 mice in each group. The mice were administered once daily for four consecutive weeks. At the end of the administration, the mice in each group were weighed, the eyeballs were taken for blood samples, and the necks were dissected and sacrificed. The livers were dissected and weighed, and the liver index was calculated. The automatic biochemical analyzer was used to detect the serum biochemical indicators of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglycerides (TG), and total cholesterol (TC) levels in mice. The ELISA method was used to detect the contents of superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT) and glutathione peroxidase (GSH-Px) in mouse 10% liver tissue homogenate. HE staining was used to observe the pathological morphology of liver tissue in mice. Results Compared with the normal group, the liver index and serum biochemical ALT, AST, TG, TC levels in the model group were significantly increased (P<0.01), and the liver homogenate MDA was significantly increased (P<0.01), while the levels of SOD, CAT, GSH- Px were decreased significantly (P<0.01). There were a large number of fatty vacuoles and hepatic cord disorders in the liver tissue. Compared with the apigenin group and the resveratrol group, the liver index, and serum biochemical ALT, AST, TG and TC levels in the combined administration group decreased (P<0.05), and the liver homogenate MDA level decreased (P<0.05), and the levels of SOD, CAT and GSH-Px were increased (P<0.05). The number of fatty vacuoles in liver tissues were reduced, hepatic cord disorders were improved. Conclusions The combined administration of apigenin and resveratrol has a protective effect on NAFLD model mice, possibly through the reduction of hepatic enzymes and blood lipid levels, as well as enhanced antioxidant activity. The combination treatment shows better efficacy compared to the apigenin and resveratrol groups.

The accumulation of basic researches and clinical studies related to cytokine-induced killer (CIK) cells has confirmed their safety and feasibility in treating malignant diseases. This review summarizes the available published literature related to the biological characteristics and clinical applications of CIK cells in recent years. A number of clinical trials with CIK cells have been implemented during the progressive phases of cancer, presenting potential widespread applications of CIK cells for the future. Furthermore, this review briefly compares clinical applications of CIK cells with those of other adoptive immunotherapeutic cells. However, at present, there are no uniform criteria or large-scale preparations of CIK cells. The overall clinical response is difficult to evaluate because of the use of autologous CIK cells. Based on these observations, several suggestions regarding uniform criteria and universal sources for CIK cell preparations and the use of CIK cells either combined with chemotherapy or alone as a primary strategy are briefly proposed in this review. Large-scale, controlled, grouped, and multi-center clinical trials on CIK cell-based immunotherapy should be conducted under strict supervision. These interventions might help to improve future clinical applications and increase the clinical curative effects of CIK cells for a broad range of malignancies in the future.

Objective To explore the feasibility of Ilizarov technique in managing large tibia defects combined with soft tissue defects.Methods A total of 24 patients with large tibial defects combined with soft tissue defects caused by compound open tibial fractures were fixed with Ilizarov technique from September 2003 to September 2010.All patients belonged to open tibial fractures,including 20 patients with Gustilo type Ⅲ B and four with Gustilo type Ⅲ C.After debridement,the soft tissue defect areas was 10 cm ×6 cm and the bone defect was(8 ±4)cm.Fifteen patients with tibial defects ＜5 cm were treated with one stage debridement,fibula resection and tibial defect end compression.The other nine patients with tibial defect ＞ 5 cm were managed by one stage debridement,bone transport and bone lengthening.Then,15 patients were treated with one stage debridement,wound closure or wound reduction,bone grafting treatment and second stage cleansing of the incarcerated skin and fracture end.Results All patients were followed up for average 14 months(10-24 months),which showed reconstruction of the bone defects,restoration of the limb length,fracture healing and less than 2 cm difference between health limb and contralateral limb.One patient experienced common peroneal nerve palsy after operation,but recovered three months later.Of all,19 patients recovered without extra surgery,three restored with skin graft and two received skin flap.Conclusion Ilizarov technique is an effective option for treating the tibial defects combined with soft tissue defects at one stage.