Objective To obtain the expression pattern of multidrug resistance-associated gene in human small cell lung cancer(SCLC)MDR cell line SH77/CDDP and provide basis for further studying the mechanism of human SCLC MDR induced by cisplatin.Methods Total RNA was isolated from SCLC MDR cell line SH77/CDDP and its parental cells,and synthesized into double-stranded cDNA,then synthesized into biotin-labeled cRNA probe by in vitro transcription.The cRNA probes were separately hybridized with Affymetrix GeneChip and human U133 set chips(U133A and U133B,containing 39 000 transcripts,including 33 000 known human genes and 6 000 unknown cDNA expression sequence tags,ESTs),and the signals were scanned by the GeneArray Scanner.The results were analyzed by bioinformatics.Results Compared with the gene expression profile of parental SH77 cells,2 389(6.13%)genes were up-regulated in SCLC MDR cell line SH77/CDDP cells,of which 461(1.19%),89(0.23%),26(0.07%)and 7(0.02%)genes were separately up-regulated one-fold,two-folds,three-folds and four-folds.36 genes,including ABCC3,HSP70,CYP26B1,CYP4A11,IGF1R,LOX2,CAP2,LRP2BP,AKNA,ELTD1 and otherwise,were up-regulated 2.5-5.1 folds.According to Gene Ontology and Tree View analysis,these 35 genes were involved in transport,cell adhesion,signal transduction,transcription and ion binding and so on.Conclusion Many multidrug resistance-associated genes induced by cisplatin have been screened by high-throughput gene chip method.Validating their cellular functions will help to identify the mechanism of human SCLC MDR induced by cisplatin.

Objective To establish a small cell lung cancer (SCLC) multidrug resistance (MDR) cell line SH77/CDDP and analyze its biological properties. Methods The SCLC MDR cell line SH77/CDDP was established by increasing gradually the dose of cisplatin, the first line chemotherapeutic drug for lung cancer. The chemosensitivities of SH77/CDDP and its parental cell line to 7 chemotherapeutic drugs were tested. Changes in cellular morphology and ultrastructure were observed. The cell growth curve and cell cycle were also determined. Results SCLC MDR cell line SH77/CDDP having different drug resistance to the 7 chemotherapeutic drugs was established successfully. Compared with that of its parental cell, the size of MDR cell was a bit bigger. The ratio of nucleus to cell plasma decreased and mitochondria, Golgi bodies, rough endoplasmic reticulums and free ribosomes increased. The finger like apophysis was obvious. The rate of cell proliferation of SH77/CDDP was similar to that of SH77, but the number of cells in S phase increased( P

Objective To study the effects of small dosage insulin on intestinal inflammatory responses to endotoxin rats. Method Thirty two male SD rats were randomly divided into 4 groups (n = 8): control group, endotoxin (LPS,6 mg/kg i.p.)group, regular insulin(RI,0.5 IU/kg hypodermic) group and LPS(6 mg/kg i.p) + RI (0.5 IU/kg hypodermic)group. Six hours after LPS or saline injection,all rats were laparotomized to observe the congestion in intestinal mucosa with naked-eyes and photography.Then a segment of intestine was stained with HE to observe the pathological changes. The expressions of IL-6 and TNF-α were detected by RT-PCR.The systemic inflammatory response,blood sugar and food taken in rats were observed simultaneously. Software SPSS 13.0 was used to perform ANOVA and Chi-square test for statistical analysis. Results Compared with LPS group, the differences in hyperemia and inflammatory cell infiltration in intestinal tissue were not noticeable in LPS + RI group. The expression of IL-6 and TNF-α were significantly attenuated in RI + LPS group (P < 0.01). All rats in LPS group manifested systemic inflammatory response syndrome (SIRS) four or five hours after LPS treatment, while there was none in LPS + RI group. Rats in LPS group took less food than rats of other groups while the blood sugar had litter difference in all groups (P > 0.05). Conclusions Small dosage of insulin could reduce intestinal inflammation caused by endotoxemia. Early administration of insulin ould prevent the presence of SIRS while it has no obvious influence on blood sugar.

Objective To design nursing occupational safety and health curriculum reasonably.Methods With a semi-structured interview outline,focus group interviews among eight clinical nursing and nursing education specialists were conducted to develop the occupational safety and health curriculum design.Results It was important to set up nursing occupational safety and health curriculum.The main object,contents,teaching and evaluation methods were identified.Conclusions Occupational safety and health curriculum can be involved in nursing curriculum system to reduce occupational hazards and maintain physical and mental health of nursing students.

Objective To study the drug sensitivity of chemotherapeutic drugs by MTT method. Methods Twenty five lung cancer specimens obtained by operation were used for primary culture and the sensitivity of the cancer cells to four drugs, adriamycin(ADM), cis dichlorodiamine platinum(DDP), vincristine(VCR) and etoposide(VP16), was tested by MTT assay. TOPOⅡ, p53, bcl 2 and GST ? proteins in 10 cases of lung cancer tissue were determined by immunohistochemistry. Results The successfully obtained results in 21 cases showed that the effective rate of ADM, DDP, VCR and VP16 was 19.0%, 38.1%, 23.8% and 23.8% respectively. Positive expressions of TOPOⅡ, p53, bcl 2 and GST ? proteins were found in lung cancer tissue in the 10 cases which had different multi drug resistance(MDR) to the four drugs. Conclusion It is possible to test the drug sensitivity of the primary cultured lung cancer cells obtained from operation by MTT method and the results are useful for the selection of anticancer drugs. The expressions of TOPOⅡ, p53, bcl 2 and GST ? are factors to determine MDR for lung cancer cells.

Objective To investigate the effect of ultrasound-guided subcostal transverses abdominis plane (TAP) block with dexmedetomidine after laparoscopic radical operation. Methods 40 patients underwent laparoscopic radical operation for colorectal cancer were randomized into dexmedetomidine group (group DEX) and control group (group CON). All the patients received ultrasound-guided subcostal TAP block after operation, Group DEX with dexmedetomidine 1 μg/kg and 0.25% ropivacaine to 20 ml, and group CON with 0.25% ropivacaine 20 ml. All the patients were assessed with Ramsay scores and the pain at rest and on coughing were assessed with Visual Analogue Scale (VAS), 2, 6, 12, 24 and 24 hours after operation. The highest level and the duration of sensory blockade, the first time and the total times of pressing the analgesia pump in the first day after operation, and the requirements of sufentanil were recorded. First flatus time, first diet time and the length of hospital stay were compared. Results The scores of VAS were significantly less (P<0.001), and the Ramsay scores were more in the group DEX than in the group CON (P<0.01) 2, 6 and 12 hours after operation; with the longer time of sensory blockade (P<0.001), the later to first press the analgesia pump (P<0.001), the less frequence of pressing the analgesia pump (P<0.001), and less dosage of sufentanil (P<0.001). The first flatus time, first diet time were significantly earlier in the group DEX than in the group CON (P<0.001), with the less length of total hospital stay (P<0.001). Conclusion Dexmedetomidine can promote the anaesthesia of ultrasound-guided subcostal TAP block with ropivacaine and improve the recovery after laparoscopic radical operation.

Objective To study the mRNA expression levels and clinical significance of omsomucoid 1-like protein 3 (ORMDL3) gene in the peripheral blood of recurrent wheeze children under 3 years old.Methods Peripheral blood specimens of 25 recurrent wheeze children including 14 non-atopy patients (group A) and 11 atopy patients (group B) that were registered in the First Affiliated Hospital of Nanjing Medical University,from Sep.2010 to Sep.2012 were enrolled based on the inclusion criteria and 24 non-allergic controls(the children with food allergy,drug allergy or ectema was excepted).The mRNA expression levels of ORMDL3 gene were detected by reverse transcription (RT)-PCR and clinical features were analyzed.Results The expression levels of ORMDL3 were up-regulated in the peripheral blood specimens of group B compared with group A (t =14.12,P ＜ 0.01).Compared with peripheral blood specimens from normal subjects,mRNA expression of ORMDL3 were significantly increased in recurrent wheeze children(t =10.29,5.73,P ＜0.01).The incidence of wheeze groups exist gender differences,male ＞ female.Wheeze usually with a high incidence in winter and spring.Conclusions The increase of ORMDL3 gene expression levels were correlated with recurrent wheeze under 3 years old especially in atopy group and may be involved in the pathogenesis of recurrent wheeze.

Objective To analyze the differential gene expression profiling of liver in rats subjected to hemorrhagic shock(HS) and sham hemorrhage shock(SHAM) by gene chip technology, thus to evaluate the possible molecular pathogenesis of HS. Method 20 male Wistar rats were randomly divided into a SHAM group and a HS group, with 10 rats in each group. Hepatic gene expression profiles were detected by oligonucleotide microarrays of 5705 mouse genes in two groups for three times. Genes with ratio(R) > 2 were identified as up-regulated and R < 0.5 were identified as down-regulated. Biological function of differentially expressed genes was analyzed and 9 genes were selected to undergo semi-quantitative RT-PCR. Results Among the total 5705 probes detected,86 genes showed differential expression in HS group comparison with SHAM group. The expression levels of 72 genes were up-regulated while those of 14 genes were down-regulated significantly. Differentially expressed genes were classified according to their biological function: transport genes, transcription regulator genes, signaling genes, response to stress genes, metabolic genes, development genes and cell adhesion genes. Conclusions cDNA microarray is an efficient and high-throughout method to survey gene expression profiles in HS.The variation of those gene expressions might be a potential pathogenic mechanism for HS that may offer a novel target for further study of therapeutic strategies of HS.

Objective To verify the targeting regulatory relationship between microRNA -939 (miR -939) and CD2 -associated protein (CD2AP).Methods The online RegRNA software was used to predict the human CD2AP promoter for potential binding sites complementary to miR -939.HEK -293T cells were cotransfected with hu-man CD2AP promoter plasmid pGL3 -2K and microRNA negative control (miR -NC)or miR -939 mimics,and the relative luciferase activity(RLA)was detected at 24 h post -transfection.HEK -293T cells were transfected with miR -NC or miR -939 mimics for 48 h,and the CD2AP mRNA expression level was detected by adopting reverse tran-script and real -time fluorescence quantification -PCR,while the CD2AP protein expression level was detected by using Western blot.Results (1 )There were 2 miR -939 binding sites at CD2AP promoter region,located at -468 to -491 and -654 to -677 upstream of initiation codon ATG (marked as +1 )relatively.(2)At 30 nmol/L,50 nmol/L,the RLA in miR -NC group and miR -939 group were 6.81 ±0.88 vs 6.07 ±2.24,5.88 ±1 .44 vs 3.94 ± 0.79 relatively,and there were no significant differences between the 2 groups (t =3.04,2.06,all P >0.05),while the RLA between the 2 groups were 5.58 ±0.58 vs 3.29 ±0.64 at 1 00 nmol/L,and the difference was significant between the 2 groups(t =4.07,P <0.05).(3)At 30 nmol/L,50 nmol/L and 100 nmol/L,the relative CD2AP mRNA expression in miR -NC group and miR -939 group were 1 .00 ±0.01 vs 0.80 ±0.08,1 .00 ±0.00 vs 0.80 ±0.1 3 and 1 .00 ± 0.00 vs 0.72 ±0.07 relatively,while the CD2AP mRNA expression was decreased by 20% -30% at each concentration level,and there were significant differences between the 2 groups (t =4.44,2.93,6.84,all P <0.05).(4)At 50 nmol/L, the relative CD2AP protein expression in miR -NC group and miR -939 group were 0.48 ±0.09 vs 0.19 ±0.12,and the CD2AP protein expression was decreased,and the difference was significant (t =3.36,P <0.05).Conclusions CD2AP is the target gene of miR -939,and miR -939 can down -regulate the expression of CD2AP both in mRNA and protein levels by targeting its promoter region,which indicates that miR -939 may mediate the podocyte injury.

Objective To investigate the indirect decompression effect of oblique lateral interbody fusion (OLIF) in the treatment of mild to moderate degenerative lumbar stenosis.Methods From October 2014 to November 2016,23 patients with mild to moderate lumbar spinal stenosis underwent OLIF combined with or without posterior pedicle screw fixation;9 males and 14 females with average age of 59.2±11.6 years old;8 cases at L3,4 segment and 15 cases at L4,5 segment.All cases were followed up for more than 6 months.Thin layer scanning of CT and two-dimensional reconstruction images were used to measure the vertical diameter and area of intervertebral foramen.Intervertebral disc height and spinal canal anteroposterior diameter were measured on median sagittal MRI sequence,and the anteroposterior diameter and the cross-sectional area of the spinal canal were measured on cross-sectional MRI sequence.The clinical effects were assessed by the visual analogue score (VAS) and the Oswestry disability index (ODI) for low back pain,lower limb pain and lower limb numbness.Results Compared with those measurements pre-operatively,the post-operative intervertebral disc height increased by 78.6％±13.4％.The post-operative left vertical diameter of intervertebral foramen increased by 36.7％±7.8％,and the post-operative left area of intervertebral foramen increased by 36.6％± 8.7％,and the post-operative right vertical diameter of intervertebral foramen increased by 40.7％±9.6％,and the post-operative right area of intervertebral foramen increased by 40.0％±8.9％.The post-operative anteroposterior diameter of sagittal spinal canal were increased 32.6％ ± 5.9％,and the post-operative anteroposterior diameter of cross-sectional spinal canal were increased 34.4％±6.8％,and the post-operative cross-sectional area of the spinal canal were increased 47.5％±7.2％.All of the differences were statistically significant between pre-operative and post-operative measurements.The VAS score for low back pain was 6.2± 1.7 pre-operatively,and 1.1±0.5 post-operatively.The ODI for low back pain was 81.2％± 18.2％ pre-operatively,and 6.1％±2.0％ post-operatively.The VAS score for lower limb pain was 5.6±1.4 pre-operatively,and 0.8±0.3 post-operatively.And the VAS score for lower limb numbness was 6.6±2.0 pre-operatively,and 3.4± 1.2 post-operatively.All of the differences were statistically significant between pre-operative and post-operative evaluations.Conclusion There were obvious radiological evidences and remarkable clinical effect of indirect decompression using OLIF technique in treatment of mild to moderate lumbar spinal stenosis at early post-operative stage.However further long-term follow-up studies with multicenter large sample were still needed.