Objective: To construct tetracycline-dependent regulatory system in one vector,which contains two multiple cloning sites(MCS) for the convenient cloning of two target genes.Methods: The tetracycline(Tet)-dependent regulatory fragment(TO)(between NdeI and SacII sites) was amplified from pcDNA4TM/TO vector with PCR,which was inserted into pUC118 vector and sequenced.The counterpart fragment in pVITRO3 was replaced with correct Tet-dependent regulatory fragment,namely pVITRO3-TO vector.The Tet repressor gene(TR) encoding the Tet repressor from pcDNA6/TR vector was incorporating into the backbone of the pVITRO3-TO.Result: The tetracycline(Tet)-dependent regulatory fragment and the Tet repressor gene were amplified successfully with PCR from its template vectors and were subcloned.The clones with correct sequence were selected by sequencing the picked 2 clones.The target vector was constructed successfully by incorporating TO and TR into the backbone of pVITRO3 vector.Conclusion: Tetracycline-dependent regulatory double-expressing system in one vector was constructed initially.

Occupational Health

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With the inappropriate use of antibiotics, the situation of bacterial resistance is more and more severe. The emergence of multidrug-resistant (MDR) bacteria has made it difficult to cure the infections in clinical. For treatment the infections caused by MDR and reducucton of the generation of resistant bacteria, researchers are actively studying on the non-antibiotic substances for antibacterial activity. In this paper, the advances in those with conifrmed effects such as phage therapy, metal/chelation therapy, immunization therapy, photodynamic therapy, and nitric oxide (NO)-based therapies, small molecule inhibitors, antimicrobial peptides and Chinese herb were reviewed.

Objective To explore the reason and influence of communication difficulties between oncology nurses and patients or families of patients,and to obtain some relevant advice about communication difficulties.Methods This study used focus group to interview 24 oncology nurses who were selected from the First Affiliated Hospital of Chongqing Medical University by homogeneous sampling method,and were randomly divided into three groups (8 in each group).The data were processed and analyzed by content analysis.Results The reasons why oncology nurses had communication difficulties with patients and their families were as follows:putting forward negative problems about their disease,ineffective communication and conflicts with patients,families.The communication difficulties showed negative effect on oncology nurses.Oncology nurses thought that the main reason of communication difficulties lied in the deficiency of institution and propounded the suggestion to resolve it.Conclusions oncology nurses were tend to have difficulty in communicating with patients and their families in the long-term work,especially in the period of crisis:early diagnosis of tumor stage,relapse,advanced cancer and some special dates (birthday,parties etc.).Therefore,oncology nurses,patients and their families need to be supported.To solve the difficulties of communication could reduce the negative emotions of patients and their family,such as anxiety,frustration,helplessness,so as to improve the quality of care and life of patients.

Hepatitis B virus(HBV)will become reactive in non-active HBsAg carriers when they are in the immunosuppression status,which is often caused by cytotoxic drugs in chemotherapy or immunosuppressants.Liver damage will be induced in such patients and even liver failure in some patients.Nucleoside analogues could be used in early phase as prophylaxis for reactivation of hepatitis B in immunosuppression and to improve clinical prognosis.

The effects of sodium salicylate on the expression of heat shock protein 27 (HSP27) during oxidative stress in tissue-cultured human lens epithelial cells were investigated. Cultured human lens epithelial cells (HLB-3) were divided into 3 groups: control group (group A), oxidation injury group (group B) and sodium salicylate group (group C). Apoptosis of human lens epithelial cells cultured in vitro was induced in the presence of 150 micromol/L H2O2. Cells viability and the expression of HSP27 were analyzed. Viability of the cells was measured by methyl thiazole tetrazolium (MTF) chromatometry. The expression of HSP27 in HLB-3 cells was detected by using immunohistochemistry and image analysis system. Sodium salicylate could induce the expression of HSP27, and the cells viability in group C was significantly higher than in group B (0.2667+/-0.01414 vs 0.2150+/-0.01080, P=0.012<0.05). The average gray value of HSP27 in group B was less than that in group C (P=0.000<0.05). The increased expression of HSP27 by sodium salicylate might play an important role in the protection of hydrogen peroxide-induced injury of human lens epithelial cells, suggesting that sodium salicylate could suppress, at least in part, the apoptosis of human lens epithelial cells.

Humans ; Occupational Diseases ; Occupational Health

Background Cell immunologic therapy for retinoblastoma(RB)is becoming a hot research topic.Cancer-testis antigen is a human immunogenic protein and is used to treat some tumors.However,its effect on RB has not been investigated.Objective The present study was to discuss the antigen specific anti-tumor effect of cytotoxic T lymphocytes(CTL)induced by the cancer-testis antigen,NY-ESO-1-sensitized dendritic cells(DCs),on human RB.Methods PCR was performed to amplify target gene fragments from the NY-ESO-1 plasmids,and then the target gene fragments were digested with the restriction enzymes SalI and EcoRI.Harvested fragments were inserted into the pDC316 plasmid to construct the recombinant plasmid pDC316/NY-ESO-1.The expression of NY-ESO-1 protein in human RB cells strain,HXO-RB44,was detected by immunofluorescence and Western blot.Monocytes were isolated from 60 ml of peripheral blood from a healthy donor using Ficoll density-gradient centrifugation with a cell density of 1 × 107/ml.DCs isolated from blood were stimulated with recombinant human granulocyte-macrophage colony stimulating factor(rhGM-CSF)and recombinant human interleukin-4(rhIL-4).The recombinant plasmid pDC316/NY-ESO-1 was transfected into DCs and the DCs were co-cultured with T lymphocytes.The resultant CTL were used as effector cells.The growth of the CTL was detected by MTT assay.The CTL were then added into the growth medium used for culturing HXO-RB44 cells and the vitality of the HXO-RB44 cells was assayed by MTT assay.Results The sequence of the cloned DNA fragment of the recombinant plasmid pDC316/NY-ESO-1 was conforms with the sequence of the NY-ESO-1 gene.The expression of the NY-ESO-1 protein in HXO-RB44 cells was tested by immunofluorescence and Western blot.DCs were successfully induced with rhIL-4 and rhGM-CSF from PBMC.The recombinant expression plasmid pDC316/NY-ESO-1 was successfully transferred into DCs.These DCs had high expression of surface molecules such as HLA-DR(42.1％),CD80(54.2％),CD83(39.7％)and CD86 (94.8％).The CTL that was induced by DCs-sensitized with NY-ESO-1 specifically killed HXO-RB44 cells.CTL induced by the sensitized DCs had a stronger cytotoxic effect against HXO-RB44 cells compared with un-sensitized DCs and CTL un-induced with DCs,as shown by MTT asssay(P＜0.05).The anti-tumor activity was highest when the ratio of effector to target was 75∶1(P＜0.05).Conclusions DCs transfected by the recombinant plasmid pDC316/NY-ESO-1 can induce the proliferation of allogenic CTLs,which showed a specific anti-tumor effect against HXO-RB44 cells.These results present a new type of immunotherapy for the treatment of RB.

Aged ; Cardiovascular Diseases ; diagnosis ; etiology ; Cerebrovascular Disorders ; diagnosis ; etiology ; Diagnostic Errors ; Female ; Humans ; Laryngitis ; complications ; diagnosis ; Middle Aged