Aim To construct the eukaryotic expression vectors of short hairpin RNA targeting survivin and observe its effect on biologic behavior of A549 cells and sensitivity of A549 cells to paclitaxel.Methods The DNA fragment targeting human survivin was inserted into the plasmid,and the recombinant plasmid was constructed.The recombinant plasmids cells were transfected into A549 cells by FuGENE transfection reagent.The expression levels of survivin gene were detected with RT-PCR and Western blot before and after transfection,respectively.Cell apoptosis was detected by TUNEL method.The sensitivity of A549 cells to paclitaxel was detected by MTT after transfection.Results The recombinant plasmid was successfully constructed.RNAi group cells showed lower expression of survivin than control group.The apoptosis rate of A549 cells increased after transfection.The IC50 of paclitaxel inhibiting A549 cells was 11.9 fold before transfection compared with those after transfection.There was significant difference between the two groups(P

Lung cancer has been the most dangerous malignant tumor in the worldwide.Nowadays,for patients with resectable early or local advanced non-small cell lung cancer (NSCLC),both domestic and foreign guidelines recommended multidisciplinary treatments like surgery,chemotherapy and radiotherapy.Recent studies have shown that neoadjuvant chemotherapy can significantly improve the prognosis of patients with resectable NSCLC and has better treatment compliance and tolerance.

OBJECTIVETo study the relationship between targeting vector structure and homologous recombination rate and investigate whether the mouse p16(INK4a) plays a role in tumor suppression.

METHODSA conditional targeting vector with 2.0 kb EcoR I/Xba I fragment as short arm and 5.9 kb SpeI/NotI fragment as long arm was built. Of the 2 direct locus crossing- over(loxPs) in the vector, one was inserted at 240 bp upstream of the initiate code of p16(INK4a) exon 1a and the other at 1633 bp downstream of the initiate code. Both exon 1a and the selection marker Neo will be deleted in targeted cells when mediated by Cre. After linearlization and purification, t he targeting vector was introduced into ES cells through electroporation.

RESULTSTwenty-four G418- and gancyclovir-resistant ES cell colonies were picked out and one of them was confirmed as positive by Southern hybridization.

CONCLUSIONTargeting vectors with 2 TK genes flanking the homologous arms are likely to produce good result of homologous recombination.

Animals ; Anti-Bacterial Agents ; pharmacology ; Antiviral Agents ; pharmacology ; Base Sequence ; Cell Division ; drug effects ; genetics ; Cell Line ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; Drug Resistance ; genetics ; Embryo, Mammalian ; cytology ; drug effects ; metabolism ; Exons ; genetics ; Ganciclovir ; pharmacology ; Genetic Vectors ; genetics ; Gentamicins ; pharmacology ; Mice ; Molecular Sequence Data ; Recombination, Genetic ; Stem Cells ; cytology ; drug effects ; metabolism ; Thymidine Kinase ; genetics ; metabolism ; Transfection

Cisplatin and nedaplatin have been proved to be effective for most of solid tumors including head and neck cancer,esophageal cancer,lung cancer,bladder cancer,ovary epithelial cancer,testicular cancer and cervical cancer. Nedaplatin is often used as an alternative to cisplatin in Asia. However,the evi-dence from phase Ⅲ clinical trials is insufficient. Clinicians should be cautious when using nedaplatin to re-place cisplatin.

Radiation pneumonitis is a common complication in the radiotherapy for thoracic malignant tumors. And the resulting respiratory failure is one of the most serious side effects. The occurrence and severity of radiation pneumonitis depend on many factors such as age, performance status scores, smoking status, lung condition, tumor sizes, tumor location, chemoradiotherapy related factors, inflammatory factors and single-nucleotide polymorphism, et al. Some imaging examinations, for example single-photon emission computed tomography and PET-CT, have been used to predict radiation pneumonitis, while its usefulness remains to be strengthened. Further researches are still needed to find the gold standard in the prediction of radiation pneumonitis.

OBJECTIVETo investigate the role of transglutaminase 3 (TGM3) gene in laryngeal carcinogenesis.

METHODSThe authors detected the deletion indirectly through loss of heterozygosity (LOH) analysis at DNA level using 4 STR primers within and near TGM3 gene in 72 cases, and detected the differential expression of TGM3 gene in 8 cases of paired normal and cancerous tissue of laryngeal carcinoma by Northern blot.

RESULTSLOH was found existing in all of the microsatellite loci, and the LOH frequencies were 25.76%, 20.00%, 38.10% and 18.75% at D20S17, D20S607, D20S99 and D20S841 respectively; LOH concerning at least one polymorphism locus accounted for 61.11%. No correlation of clinical stage, lymph node metastasis and differentiation with the LOH of TGM3 gene was observed, P>0.05. TGM3 gene expressed significantly higher in normal tissues than in paired cancerous tissues.

CONCLUSIONTGM3 gene might play an important role in laryngeal carcinogenesis and further researches will be needed to clarify the possible mechanisms.

Blotting, Northern ; Calcium-Binding Proteins ; genetics ; DNA, Neoplasm ; genetics ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Neoplastic ; Humans ; Laryngeal Neoplasms ; enzymology ; genetics ; Loss of Heterozygosity ; Microsatellite Repeats ; RNA, Neoplasm ; genetics ; metabolism ; Transglutaminases ; genetics

Objective    To evaluate the clinical effects of segmentectomy versus lobectomy under single utility port video-assisted thoracic surgery on inflammatory factors and immune cells in peripheral blood of non-small cell lung cancer patients, and to analyze the effect of changes of postoperative inflammatory factors and immune cells on the prognosis of the patients. Methods    The clinical data of 256 patients who underwent segmentectomy or lobectomy under single utility port video-assisted thoracic surgery for non-small cell lung cancer in the First Affiliated Hospital of Hebei North University from January 2016 to October 2020 were retrospectively collected. According to the operation method, they were divided into a segmentectomy group (126 patients with 79 males and 47 females at an age of 63.4±6.2 years) and a lobectomy group (130 patients with 91 males and 39 females at an age of 62.9±5.6 years). The change of inflammatory factors (C reactive protein, interleukin-6, interleukin-8, tumor necrosis factor-α) and immune cells (CD4+T cells, CD8+T cells and natural killer cells) were recorded and analyzed before operation (T0) and 1 day (T1), 3 days (T2), 7 days (T3), 1 month (T4) after the operation between the two groups. According to postoperative recurrence situations, they were divided into a recurrence group and a non-recurrence group, multivariate logistic regression analysis was used to analyze the relationship between the change of postoperative inflammatory factors, immune cells, and the prognosis of patients with non-small cell lung cancer. Results    (1) There was no statistical difference in sex ratio, underlying diseases, body mass index, levels of preoperative inflammatory factors or immune cells between the two groups (all P>0.05). (2) The changes of postoperative inflammatory factors in the segmentectomy group were significantly less than those in the lobectomy group at T1-T3 (all P<0.05), and the changes of postoperative immune cells in the segmentectomy group were significantly less than those in the lobectomy group at T1-T4 (all P<0.05). (3) The changes of postoperative inflammatory factors and immune cells on postoperative day 3 in the recurrence group were significantly more than those in the non-recurrence group (all P<0.05). (4) Multivariate logistic regression analysis showed that the changes of postoperative inflammatory factors and immune cells on postoperative day 3 may be the risk factors for postoperative recurrence and metastasis in patients with non-small cell lung cancer (all P<0.05). Conclusion    Single utility port video-assisted thoracic surgery segmentectomy for the treatment of non-small cell lung cancer can reduce the inflammatory response and protect body's immune function, and the change of postoperative inflammatory factors and immune cells in postoperative day 3 may be the risk factors for postoperative recurrence and metastasis in patients with non-small cell lung cancer.