Overexpression of cyclooxygenase-2 in human HepG2, Bel-7402 and SMMC-7721 hepatoma cell lines and mechanism of cyclooxygenase-2 selective inhibitor celecoxib-induced cell growth inhibition and apoptosis

AIM： To investigate the cyclooxygenase-2 （COX-2） expression level in human HepG2, Bel-7402 and SMMC-7721 hepatoma cell lines and the molecular mechanism of COX-2 selective inhibitor celecoxib-induced cell growth inhibition and cell apoptosis. METHODS： Hepatoma cells were cultured and treated with celecoxib. Cell in situ hybridization （ISH） and immunocytochemistry were used to detect COX-2 mRNA and protein expression. Proliferating cell nuclear antigen and phosphorylated Akt were also detected by immunocytochemistry assay. Cell growth rates were assessed by 3-（4, 5-dimethylthiazol-2-yl-2, 5-diphenyltetrazolium （MTT） bromide colorimetric assay. Celecoxib- induced cell apoptosis was measured by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） and flow cytometry （FCM）. The phosphorylated Akt and activated fragments of caspase-9, caspase-3 were examined by Western blotting analysis. RESULTS： Increased COX-2 mRNA and protein expression were detected in all three hepatoma cell lines. Celecoxib could significantly inhibit cell growth and the inhibitory effect was in a dose- and time-dependent manner evidenced by MTT assays and morphological changes. The apoptotic index measured by TUNEL increased correspondingly with the increased concentration of celecoxib and the reaction time. With 50 μmol/L celecoxib treatment for 24 h, the apoptotic index of HepG2, BEL-7402 and SMMC-7721 cells was 25.01±3.08%, 26.40±3.05%,and 30.60±2.89%, respectively. Western blotting analysis showed remarkable activation of caspase-9, caspase-3 and dephosphorylation of Akt （Thr^308）. Immunocytochemistry also showed the reduction of PCNA expression and phosphorylation Akt （Thr^308） after treatment with celecoxib. CONCLUSION： COX-2 mRNA and protein overexpression in HepG2, Bel-7402 and SMMC-7721 cell lines correlate with the increased cell growth rate. Celecoxib can inhibit proliferation and induce apoptosis of hepatoma cell strains in a dose- and time-dependent manner.

Current applications and future prospects of nanotechnology in cancer immunotherapy

Cancer immunotherapy is an artificial stimulation of the immune system to recognize cancer cells and activate specific immune cells to target and attack cancer cells.In clinical trials, immunotherapy has recently shown impressive results in the treatment of multiple cancers.Thus, cancer immunotherapy has gained a lot of attention for its unique advantages and promising future.With extensive research on cancer immunotherapy, its safety and effectiveness has gradually been revealed.However, it is still a huge challenge to expand and drive this therapy while maintaining low toxicity, high specificity, and long-lasting efficacy.As a unique technology, nanotechnology has been applied in many fields, the advantages of which will promote the development of cancer immunotherapies.Researchers have tried to apply nanomaterials to cancer immunotherapy due to their advantageous properties,such as large specific surface areas, effective drug delivery, and controlled surface chemistry, to improve treatment efficacy.Here,we briefly introduce the current applications of nanomaterials in cancer immunotherapy, including adoptive cell therapy(ACT),therapeutic cancer vaccines, and monoclonal antibodies, and throw light on future directions of nanotechnology-based cancer immunotherapy.

Magnetically labeled mesenchymal stem cells after autologous transplantation into acutely injured liver

AIM:To evaluate tracking of magnetically labeled mesenchymal stem cells(MSCs) after intraportal transplantation.METHODS:Mononuclear cells were isolated from bone marrow aspirates of pigs by density gradient centrifugation,cultured and expanded,after which,they were incubated with super paramagnetic iron oxide(SPIO).Prussian blue staining was performed to highlight intracellular iron.To establish swine models of acute liver injury,0.5 g/kg D-galactosamine was administrated to 10 pigs,six of which were injected via their portal veins with SPIO-labeled MSCs,while the remaining four were injected with unlabeled cells.Magnetic resonance imaging(MRI) was performed with a clinical 1.5T MR scanner immediately before transplantation and 6 h,3 d,7 d and 14 d after transplantation.Prussian blue staining was again performed with the tissue slices at the endpoint.RESULTS:Prussian blue staining of SPIO-labeled MSCs had a labeling efficiency of almost 100%.Signal intensity loss in the liver by SPIO labeling on the FFE(T2*WI) sequence persisted until 14 d after transplantation.Histological analysis by Prussian blue staining confirmed homing of labeled MSCs in the liver after 14 d;primarily distributed in hepatic sinusoids and liver parenchyma.CONCLUSION:MSCs were successfully labeled with SPIO in vitro.MRI can monitor magnetically labeled MSCs transplanted into the liver.

Randomized clinical trial on seven-day-per-week continuous accelerated irradiation for patients with esophageal carcinoma: Preliminary report on tumor response and acute toxicity

AIM: Tumor response and normal tissue toxicity of seven-day-per-week continuous accelerated irradiation (CAIR) for patients with esophageal carcinoma were evaluated and compared to conventional irradiation (CR). METHODS: Sixty patients with squamous cell carcinoma of the esophagus were randomized into two groups: the CAIR group (30 patients) and the CR group (30 pa- tients). Patients in the CAIR group received radiotherapy (RT) with 2 Gy/fraction per day at 7 d/wk with a total dose of 50-70 Gy (average dose 64.2 Gy). The overall time of irradiation was 3.6-5.0 wk (average 4.6 wk). RT in the CR group was 2 Gy/fraction per day at 5 d/wk with a total dose of 40-70 Gy (average dose 61.7 Gy). The overall time of irradiation was 4.0-7.0 wk (average 6.4 wk). RESULTS: The data showed that the immediate tumor response to RT was better in the CAIR group than in the CR group. Efficiency rates (CR plus PR) were 82.8% (24/29) and 58.6% (17/29), respectively (P = 0.047). In both groups the incidences of esophagitis and tracheitis were insignificant (P = 0.376, 0.959), and no patient re- ceived toxicity that could not be tolerated. CONCLUSION: CAIR shortens overall treatment time and is well tolerated by patients. It may be superior to CR in enhancing the local response of tumor, but its remote effect for esophageal carcinoma awaits further follow-up.

Clinical study about endoscopic inguinal lymphadenectomy for patients with vulvar carcinoma

Objective： The aim of this study was to explore the new method of inguinal lymphadenectomy in order to reduce side effects of conventional method for patients with vulvar carcinoma. Methods： Lipolysis and liposuction were performed to subcutaneous fat on inguinal region. We inserted endoscope and filled with C02 gases to this field and then resected inguinal lymph nodes with ultrasonic scalpel. The operative field was placed with the vacuum sealing drainage and pressured with soft saline bag after the operation. Results： Many lymphatic vessel, small blood vessels and hanging lymph nodes in the subcutaneous tissues of inguinal region were revealed after lipolysis and liposuction and lymph nodes can be easily removed. The follow-up so far showed that healing of the incision was good and there was no lymphedema of patient＇s lower limb and inguinal region. Conclusion： Endoscopic inguinal lymphadenectomy can resect the lymph nodes and keep most of the lym- phatic vessels. So this technique has less influence on the lymph backflow of lower limb and inguinal region and can avoid the huge incision of conventional method. This method is worthy of further study.

Effect of ginseng polysaccharides and dendritic cells on the balance of Th1/Th2 T helper cells in patients with non-small cell lung cancer

OBJECTIVE: To investigate the effect of thorascopic administration of ginseng polysaccharides(GPS)plus dendritic cells(DC) on T helper cell type 1/T helper cell type 2(Th1/Th2) balance in patients with non-small cell lung cancer(NSCLC).METHODS: A total of 96 NSCLC patients were divided evenly into two groups. The control group was treated with DCs alone and the treatment group was treated with DCs plus GPS. After DCs and GPS were administered thoracoscopically, once a week,4 times for 30 days, the patients' quality of life was measured with the Functional Assessment of Can-cer Treatment-Lung(FACT-L) questionnaire before and after treatment. Serum interferon-γ(INF-γ), interleukin-4(IL-4), IL-2 and IL-5 were examined before and after treatments.RESULTS: The level of Th1 cytokines(INF-γ, IL-2)and the ratio of Th1/Th2 cytokines(INF-γ/IL-4, IL-2/IL-5) increased in both treatment groups, while Th2cytokines(IL-4, IL-5) and FACT-L scores decreased(P<0.01). Furthermore, after treatment Th1 cytokines(INF-γ, IL-2) and the ratio of Th1/Th2 cytokines(INF-γ/IL-4, IL-2/IL-5) were higher in the DCs +GPS group than in the control group(P<0.05). Conversely, FACT-L scores and Th2 cytokines(IL-4, IL-5)were higher in the control group than in the DCs +GPS group(P<0.05).CONCLUSION: The treatment regime of DCs plus GPS had a greater effect on NSCLC patients' immune function as compared with DCs alone. This was evident by increased expression of Th1 cytokines(INF-γ, IL-2) and the ratio of Th1/Th2(INF-γ/IL-4, IL-2/IL-5), as well as by decreased FACT-L scores and the expression of Th2 cytokines(IL-4,IL-5).

DK crush technique: modified treatment of bifurcation lesions in coronary artery

Bifurcation lesions are still technically challenging even in the era of modern stents. High incidence of restenosis both in main vessel and side branch limits the long-term prognosis although several kinds of techniques have been identified to be successful for coronary bifurcations. Reports have demonstrated the main reason for higher incidence of ostial side branch even though drugeluting stent used in side vessel lies in that there were gaps in metal coverage and drug application. Therefore,

EBV LMP2A-specific T Cell Immune Responses Elicited by Dendritic Cells Loaded with LMP2A Protein

Type Ⅱ Epstein-Barr virus （EBV） associated malignancies such as nasopharyngeal carcinoma and non-Hodgkin＇s lymphomas consistently express latent membrane 2A （LMP2A） proteins, which have been suggested to be an ideal target for immunotherapy. In previous studies we have demonstrated that using LMP2A protein loaded dendritic cells, the most powerful antigen processing cells in the body can elicit specific and robust anti-tumor cellular immune response in vitro. In this paper, we further investigated the T cell profile of the anti-tumor immune response. We found that LMP2A specific CD4＋ and CD8＋ T cells could be stimulated by LMP2A protein loaded dendritic cells （DCs）. The Thl type immune response is dominant in the immune response mediated by LMP2A specific CD4^＋ T cells. The CD8^＋ cytotoxic T cells can lyse LMP2A bearing cells effectively and specifically. The CD8^＋ cytotoxic T cells can also secrete high level of intracellular IFN-γ, which indicates these cells are EBV-LMP2A specific cytotoxic T cells. Altogether, our studies proved that LMP2A protein loaded DCs can elicit anti-tumor cellular immune responses efficiently. This study provides a rationale for the DC-based immunotherapy against EBV-LMP2A expressing malignancies.

Association between genetic variants of the leukotriene biosynthesis pathway and the risk of stroke： a case-control study in the Chinese Han population

Background Leukotrienes are arachidonic acid derivatives long known for their inflammatory properties. Leukotriene- based inflammation has been demonstrated to play a crucial role in atherosclerosis, a major risk factor for several human diseases. Recently, human genetic studies from us and others suggest that single nucleotide polymorphisms （SNPs） in leukotriene pathway genes influence the risk of atherosclerotic diseases such as stroke. This study aimed to assess the role of additional leukotriene pathway genes as a stroke risk factor within the Chinese Hart population. Methods We sequenced the promoter, exonic, and intronic regions of leukotriene A4 hydrolase （LTA4H） and arachidonate 5-1ipoxygenase （ALOX5）, and then genotyped five SNPs in LTA#H and four SNPs in ALOX5 among 691 cases with stroke and 732 controls from the Chinese population. Results We detected a significant association between an intronic SNP in LTA4H （rs6538697） and stroke in our subjects （adjusted odds ratio, recessive model, 1.75; P=0.022）; and the SNP rs2029253 in ALOX5 was associated with a decreased risk of stroke （adjusted odds ratio, 0.76; 95% confidence interval, 0.59-0.97）. Conclusion Genetic variants in LTA4H and ALOX5 may modulate the risk of stroke in the Chinese Han population.

Midterm outcomes of percutaneous transluminal septal myocardial ablation in patients with hypertrophic obstructive cardiomyopathy refractory to medication

Hypertrophic obstructive cardiomyopathy （HOCM） is a genetic disorder characterized by severe asymmetric hypertrophy of the interventricular septum （IVS） in the absence of any other systemic or cardiac diseases. The predominant abnormal haemodynamics are caused by increased left ventricular outflow tract pressure gradient （LVOTG） and abnormal systolic anterior motion of mitral valve.

Polymorphisms of dihydropyrimidine dehydrogenase gene and clinical outcomes of gastric cancer patients treated with fluorouracil-based adjuvant chemotherapy in Chinese population

Background Dihydropyrimidine dehydrogenase （DPD）, a key enzyme involved in the catabolism of 5-fluorouracil （5-FU）, is the attractive candidate for pharmacogenetic research on efficacies and toxicities of 5-FU. The aim of this study is to explore the association between polymorphisms of dihydropyrimidine dehydrogenase gene （DPYD） and clinical outcomes of gastric cancer patients treated with fluorouracil-based adjuvant chemotherapy in the Chinese population.

Downregulation of lncRNA-HOXA11-AS modulates proliferation and stemness in Glioma cells

Background: Glioma stem cells (GSCs) represent a subpopulation of cells within glioma that are characterized by chemotherapy resistance and tumor recurrence. GSCs are therefore important therapeutic target for glioma therapy.Long non-coding RNAs (lncRNAs) have been shown to regulate important functions in cancer. HOXA11-AS is one such lncRNA and has been shown to regulate cell proliferation via promotion of cell cycle progression in glioblastoma (GBM) cells. However, the specific roles of HOXA11-AS in GSCs remain unclear.Methods: Here we investigated the role of HOXA11-AS in driving GSC stemness properties via sphere-forming and protein chip assays.Results: Gain-of-function as well as loss-of-function results showed that the HOXA11-AS maybe a critical modulator in GBM recurrence as demonstrated by cell sphere-forming ability. Furthermore, we showed that induced expression of HOXA11-AS does increase the levels of stemness-related transcription factors (Oct4/Sox17/Sox2) in U87MG cells. In vivo xenograft experiments using the HOXA11-AS knockdown U87MG cells revealed that downregulation of HOXA11-AS could strongly inhibit tumor growth. Furthermore, we found that HOXA11-AS knockdown decreased the expression of cancer stemness markers in vivo.Conclusions: Collectively, these data suggests that HOXA11-AS is involved in GSC stemness and supports its clinical significance as a important therapeutic target in glioma.

Polymorphism of methylenetetrahydrofolate reductase gene is associated with response to fluorouracil-based chemotherapy in Chinese patients with gastric cancer

Background The importance of polymorphisms in the methylenetetrahydrofolate reductase （MTHFR） gene for the prediction of the response to fluorouracil-based adjuvant chemotherapy in gastric cancer patients remains unclear. The aim of this study is to assess the predictive value of several polymorphisms of the MTHFR gene for clinical outcomes of gastric cancer patients treated with fluorouracil-based adjuvant chemotherapy in Chinese population. Methods Three hundred and sixty-two Chinese patients with gastric cancer were treated with fluorouracil-based adjuvant chemotherapy. DNA samples were isolated from peripheral blood collected before treatment. The three single nucleotide polymorphisms （SNPs） （rs1801131, rs1801133, rs2274976） genotypes of the MTHFR gene were determined by matrix- assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF MS）. Results The average response rate for chemotherapy was 46.7%. Homozygous genotypes rs2274976G/G （X2=22.7, P 〈0.01） and rs1801131A/A （X2=14.3, P=0.008） were over-represented in responsive patients. Carriers of the rs2274976A allele genotypes （G/A and A/A） and of the rs1801131C allele genotypes （A/C and C/C）were prevalent in nonresponsive patients. In the haplotype association analysis, there was a significant difference in global haplotype distribution between the groups （X2=20.69, P=0.000 124）. Conclusions These results suggest that polymorphisms of the MTHFR gene may be used as predictors of the response to fluorouracil-based chemotherapy for gastric cancer patients in Chinese population. Well-designed, comprehensive, and prospective studies on determining these polymorphisms of MTHFR gene as clinical markers for predicting the response to fluorouracil-based therapy in gastric cancer patients is warranted.