Objective To investigate the changes in cells on cell cycle,cell apoptosis and susceptibility to chemotherapeutic drugs of lung adenocarcinoma A549 cells under two dimensional (2D system) and three dimensional (3D system) culture conditions. Methods The three dimensional culture of A549 cells was carried out using rotatable culture bottle and desk-top water bath shake. Flow cytometer,in situ cell apoptosis detection kit,MTT and blood cell counter were employed to compare cell cycle,cell apoptosis and susceptibility to ADM of A549 cells under different culture conditions. Results The three dimensional culture system was confirmed to have been successful by the aid of observation under inverted and electron microscope. There were significant retardation of G_ 1 phase,lower cell apoptosis rate and decreased susceptibility to ADM in A549 cells in 3D system compared with those in 2D system. Conclusions There were remarkable differences in biological characteristics of A549 cells in two culture systems,indicating that A549 cells cultivated in 3D system simulated better solid tumor in vivo . The 3D system was very useful for further investigation of the behavior of solid tumor,so that anti-carcinoma chemotherapeutic drugs could be advantageously tested in vitro before clinical application.

Objective To find an effective method for the treatment of mitomycin C extravasation injuries. Methods The rabbit model of mitomycin C extravasation was made and managed by different methods. The therapeutic efficacy in each group was assessed by using regression index and regression time. Results The therapeutic efficacy in ice compress group and ice compress plus amifostine group was better than that in other groups and the regression time was the shortest. There was no inflammation and necrosis in skin in ice compress group. Conclusion Ice compress, which can prevent inflammation and necrosis in skin, is one of the most effective treatment methods for mitomycin C extravasation injuries.

Objective To evaluate the efficacy and toxicity of the combination of gemcitabine and cisplatin in the treatment of patients with advanced non-small cell lung cancer (NSCLC). Methods A total of 30 cases of non-small cell lung cancer (stages Ⅲ and Ⅳ) were treated with gemcitabine (1 200 mg/m 2, iv infusion, at 1 and 8 d) and cisplatin (100 mg/m 2, iv infusion, at 1 d or cisplatin at 30 mg/m 2, iv infusion, at 1 and 8 d). The administration course was 28 d. Results An objective response was obtained in 46.67% of patients (2 complete and 12 partial responses), whereas 11 patients had no change and 5 patients were progressive. The response rate was 52.38% in patients with no prior chemotherapy and the response rate of 33.33% was achieved in patients who had been given prior treatment. There was significant difference between the two groups (P

Objective To study the mechanism of multicellular drug resistance of pulmonary adenocarcinoma cell line A549 mediated by cell-cell adhesion. Methods We compared the sensitivity of monolayer cells (MCs) to adriamycin (ADM) with that of multicellular spheroids (MCSs) which was employed as a three dimensional cell culture model. Transmission electron microscopy was applied to observe the cellular ultrastructure. Bcl-2, Bcl-xL, and Bax expression levels were detected by flow cytometry and indirect immunofluorescent staining. Results Compared with MCs, MCSs had more than two layers of cells, more extensive and compact cell adhesion, and inlay junctions were found within them. MCSs were more resistant to ADM. At the same time, Bcl-2 and Bcl-xL expressions in MCSs were much higher. After treatment with ADM, expressions Bcl-2 and Bcl-xL increased markedly in MCSs. Conclusion MCSs could simulate the solid tumors in vivo and has multicellular drug resistance mediated by cell-cell adhesion. The possible mechanisms may be associated with the upregulation of Bcl-2 and Bcl-xL.

OBJECTIVE To design molecular beacon probe of 43 Codon of high mutation site in rpsL of STR resistant MTB and its amplification system, meanwhile, to find the way of detecting the fluorescent light and making a qualitative judgment by use of fluorescence microscope and image analysis software. METHODS The software, Beacon designer, was used to design the 43 Codon molecular beacon probe and the amplification system, and the fluorescence microscope and image analysis software were used to detect the fluorescent light and make a qualitative judgment. RESULTS The strap of amplification product was shown clearly. The difference between PCR products from standard strain and STR resistant one was obvious in detecting the fluorescent light by use of fluorescence microscope. The rate of resistant STR detected was about 80% (P

Objective To investigate how patients′ suicide events affects the nurses psychologically ,and seeking for the proper intervention when it happens .Methods Using the Zung self‐evaluating forms (SAS)and self‐made questionnaire to investi‐gate 41 oncology nurse and analyze the data .Results Three days after the patients′ suicide events ,the nurses′ SAS score was (63 .30 ± 9 .21) ,which was prominently different from the average score of nurses who did not face incidents like this (P< 0 .05) ;during the following four weeks ,the nurses′ working state and personal life experience was hugely influenced ,experience of mental stress were severe .Conclusion The management should realize that the impact ,which was caused by these incidents that the pa‐tients committed suicide ,would render psychological damage to the nurses .Therefore ,it is necessary to build up an intervention sys‐tem to prevent the nurses from suffering psychological problems .

Objective To explore an optimal model of three dimensional in vitro cell culture for simulating solid tumors in vivo . Methods The model of three dimensional cell culture was constructed under the conditions of inhibiting the cell wall attachment and stirring the medium. Multicellular spheroids (MCS) were cultured using microcarriers (CutiSpher). Drug sensitivity of monolayer cells (MC) and MCS was tested by MTT staining and cytometry, respectively. Ultrastructures of the MC and MCS were observed by transmission electron microscopy. Results Cells in three dimensional cell culture model without microcarriers were compacted into mass at 4 d while cells in MCS were found to attach to the microcarriers at 0.5 h. MCS had more than two layers of cells growing within it at 5 d. Compared with MC, MCS was more resistant to the anticancer drug, and had more plenty of organell and microvilli with more extensive and compact cell adhesion. Conclusion MCS has strong developmental properties and can simulate the cell cell interactions in vivo , especially cell adhesion, which may contribute to the drug resistance of MCS.

Objective To construct Neuropilins-2 eukaryotic expression vector for RNA interference.Methods Recombinant targeting on gene NRP2 was designed and established with plasmid pGenSil-1 based on NRP2 cDNA equences of Genomes.Two pairs of oligonucleotides were synthesized according to the Tuschl and inserted into plasmid pGenSil-l to generate siRNA eukaryotic expression vector,DH5? strains were transformed,plasmid were extracted,and recombinant vectors were identified by the restriction map and the sequence analysis.The recombinant plasmid(pGenSil-NRP2) was transfected into the cultured LOVO cells.At 48 h after transfection,the whole cell protein was extracted,and the protein level was detected by Western blotting with mouse-anti-human NRP2 monoclonal antibody.Results Recombinant plasmids were completely coincided with the designs by the restriction map and the sequence analysis.pGenSil-NRP2 expression vector into LOVO cells down-regulated the protein level of NRP2 at 48 h after transfection.The recombinant eukaryotic expression vector were constructed successfully.Conclusion siRNA recombinant can be constructed successfully by RNAi technique for inhibiting NRP2 expression.

BACKGROUNDNo chemotherapeutic regimen and agent are effective for most patients with advanced non-small cell lung cancer (NSCLC). This study is designed to evaluate the efficacy and toxicity of the combination of gemcitabine and cisplatin in the treatment of NSCLC..

METHODSSixty cases of NSCLC in stage III and IV were treated with gemcitabine 1200mg/m² by intravenous infusion on 1st and 8th days, and cisplatin 100mg/m² by intravenous infusion on 1st day or 30mg/m² by intravenous infusion on 1st and 8th days in a 28-day cycle. Each patient was treated at least for 2 cycles.

RESULTSAn objective response was obtained in 46.67% of patients (3 complete and 25 partial responses), whereas 22 patients had no change and 10 patients were progressive. The response rate was 57.14% in patients without prior chemotherapy and 22.22% in patients with prior treatment. Significant difference existed between the two groups (P < 0.05). The main toxicities were leukopenia and thrombocytopenia, but didn't influence the chemotherapy.

CONCLUSIONSThe combination of gemcitabine and cisplatin is a feasible, well-tolerated and effective scheme in the treatment of advanced NSCLC.

Objective:To evaluate the safety and efficacy of enhanced recovery after surgery(ERAS) in the perioperative period of congenital choledochal cysts in children.Methods:This is a multicenter prospective randomized controlled study. The clinical data of 273 pediatric congenital choledochal cysts(CCC) patients who underwent surgery at 14 medical centers with complete follow-up data were collected through the medical data analysis platform. Among them, 123 cases in ERAS group were managed perioperatively in strict accordance with ERAS mode, and 150 cases in conventional group were managed according to traditional mode. The length of hospital stay,time to first farting, time to complete feeding, the incidence of complications, cost and readmission rate within 30 days,stress indexes and liver function were compared between the two groups.Results:Compared with the conventional group, median time to start farting (2.0 d vs. 3.0 d, P<0.001), median time to complete feeding (5.0 d vs. 7.0 d, P<0.001), median postoperative hospitalization time (6.0 d vs. 9.0 d, P<0.001),the median total length of stay(13.0 d vs. 15.0 d, P<0.001) were shorter,the median hospitalization cost (37,000 yuan vs.43,000 yuan P<0.001) was lower, and stress indexes recovered quickly. The incidence of postoperative hospital stay and readimission rate within 30 d were not statistically different between the two groups. Conclusion:It is safe and feasible to implement ERAS for children with CCC in the perioperative period, which can reduce stress response, speed up recovery,and save medical costs.