Objective To investigate the possibility and accuracy of idenfication of the sentinel lymph node(SLN) in breast cancer(BC) by using methylene blue intra operatively.Methods 4～6ml methylene blue dye was injected around the breast mass intra operatively. 5～10 minutes after injection,the operation for BC was performed. All blue stained lymph nodes found during operation were considered as SLN, and removed for pathological examination. All patients had axillary lymphadenectomy(ALDN),and the acquired nonsentinel lymph nodes(NSLN)from them were also examined. Results The SLN were identified in 45 of 50 patients(90.0%).There were all 117 SLNs in this series. Of 117 SLNs,111 were located in stage I lymph nodes in axilla(95.0%),and 6 SLNs in stage II(5.0%). Of the 5 cases of the unidentified SLN,the primary tumor was located in the inferior quadrant in 4 cases,and in the uper quadrant in 1 case (P0.05). Histologic status of the SLNs accurately predicted axillary nodes status with an accuracy rate of 91.0%. 4 cases or SLN examination were negative while their NSLN were positive with false negative rate 8.9%. Conclusions This technique can identify SLN accurately;the histology of SLN can predict axillary status correctly. Sentinel lymph node has correlation with the location of the primary tumor but has no correlation with the size of the tumor or breast operation before.

Objective To explore the teaching effect of sandwich teaching in medical psychol-ogy. Methods Totally 101 preventive medicine majors of five-year program were divided into two groups:control group(n=52) and experimental group(n=49). Students in control group were taught by traditional teaching while those in experimental group by sandwich teaching method. Teaching qualities were evaluated by final exam and questionnaire. Exam scores were expressed as x±s and were ana-lyzed by t test. Questionnaine results were expressed as number of people and percentage. Results Scores of experimental group in multiplechoice, case study and final grade (30.347±4.171, 32.031± 2.781, 74.296±5.642, respectively) were better than those of control group (28.520±4.443, 28.760± 4.305, 70.010±7.783, respectively), with significant differences between two groups(P<0.05 for all). But there was no significant difference between two groups in short-answer questions score (11.918± 2.431, 12.731±2.523, respectively)(P>0.05). More than 80.0% students in experimental group thought sandwich teaching improved their communication collaborative capacity , logical thinking ability and problem solving ability, etc. Conclusions With satisfactory teaching effect, sandwich teaching promotes teaching quality.

Background:Gemcitabine is the first-line drug for chemotherapy of pancreatic cancer. However,owing to the inherent and acquired resistance,gemcitabine does not change obviously the prognosis of patients with pancreatic cancer. Exploration of the mechanism of acquired resistance to gemcitabine is of great clinical importance. Aims:To establish a gemcitabine-resistant human pancreatic cancer cell subclone and to explore preliminarily the resistance mechanism. Methods:Human pancreatic cancer cell line SW1990 was stimulated continuously with 0. 5 μmol/ L gemcitabine in vitro to establish the gemcitabine-resistant subclone SW1990-0. 5. The resistance index of SW1990-0. 5 cells was counted by CCK-8 assay. Proliferation and invasion of SW1990 and SW1990-0. 5 cells were detected by cell doubling time assay and scratch wound healing assay in vitro;cell cycle and cell apoptosis were detected by flow cytometry;expressions of multidrug-resistance related genes(MDR-1,MRP-1,and BRCP)and gemcitabine metabolic enzyme related genes(dCK,RRM1, and RRM2)were determined by real-time PCR. Results:The resistance index of SW1990-0. 5 cells was 9. 32. Compared with the parental SW1990 cells,the proliferation capacity but not the invasion capacity of SW1990-0. 5 cells in vitro was reduced. When treated with gemcitabine,the cell cycle of SW1990-0. 5 cells was similar to that of parental cells,whereas the cell apoptosis was significantly inhibited;expressions of MRP-1,BRCP and dCK mRNA were down-regulated,while expressions of MDR-1,RRM1 and RRM2 mRNA did not change. Conclusions:A stable gemcitabine-resistant human pancreatic cancer cell subclone SW1990-0. 5 was successfully established. Inhibition of cell apoptosis and down-regulation of dCK expression might contribute to the acquired resistance to gemcitabine of pancreatic cancer.

Objective:To study the effects of a potassium channel blocker 4-Amino pyridine(4-AP)on the proliferation of human tongue squamous cell carcinoma Tca8113 cells.Methods:CCK-8 assay was used to detect the proliferation of Tca8113 cells cultured with 4-AP at the concentration of 1,5,10,20,50 and 100 mmol/L for 12,24 and 48 h respectively,the cell proliferation inhibition rate was calculated,the cell cycle distribution was examined by flow cytometry.Data were analyzed by SPSS19.0 software.Results:With the increase of 4-AP concentration and culture time,cells showed some morphologic changes.4-AP at 5 -100 mmol/L dose and time dependently inhibited the proliferation,with 24 h exposure dose dependenty decreased S-phase population and increased G0 /G1 phase population of Tca8113 cells.Conclusion:4-AP may inhibit Tca8113 cell proliferation by regulation of the cell cycle distribution.

A new method based on the multiple locus variable number tandem repeat analysis (MLVA) was applied for the genotyping of combined HIV Mycobacterium tuberculosis in Dali to investigate the genotyping and distribution pattern of Myco‐bacterium tuberculosis clinical isolates with MLVA .Mycobacterium tuberculosis clinical isolates were selected from Dali area , and the polymorphism of VNTR locus was tested with PCR .The clustering of genotype was analyzed by BioNumerics (6 .6) . Result showed that 15 VNTR loci of 61 combined HIV Mycobacterium tuberculosis clinical isolates were analyzed respectively . There were obvious polymorphisms of VNTRs .The discrimination power of these loci appeared different from each other ,with the biggest Hunter‐Gaston index (0 .839) loci was MIRU26 ,and the smallest one (0 .341) loci was MIRU4 .The clustering of genotype showed that these strains could be categorized into 5 gene clusters and 61 genotype ,the proportions of cluster Ⅰ was the biggest one ,51 .6% were cluster Ⅰ which including 32 strains .The standard strain H37Rv was belongs to cluster Ⅱ .Its indicated that there are obvious polymorphisms of VNTRs of combined HIV Mycobacterium tuberculosis clinical isolates in Da‐li .The main genotype was Beijing family genotype .

Objective To detect the expression of cyclooxygenase(COX) in human gastric cancer cell lines and its subcellular location of the isoforms. Methods Immunohistochemistry、RTPCR combined with laser scanning cofocal microscopy (LSCM) were used to investigate expression and distribution of COX. Results Positive staining of COX2 and COX1 protein was seen in human gastric cancer cell line MKN45、SGC7901 and AGS. However, the COX2 staining was absent and COX1 staining was weak in MGC803; though their mRNA in all the four cell lines. When compared to COX1,COX2 showed a stronger signal at both protein and mRNA levels of the gastric cancer cell lines, which was confirmed by double labeling and LSCM. The quantitative analysis of fluorescein intensity indicated the pixel intensity peak of COX2 reached 50～70, while COX1 only 10. LSCM also showed that COX2 was both cytoplasmic and nuclear envelope staining but COX1 only in cytoplasm. Conclusions In human gastric cancer, there is stronger expression of COX2 than COX1, and different distribution of the two isoforms implies their distinct roles in cell function.

OBJECTIVE To investigate whether efflux mechanism is involved in fluoroquinolone-resistant Klebsiella pneumoniae strains isolated in China. METHODS We compared the ciprofloxacin accumulation in clinically isolated K. pneumoniae with or without CCCP by fluorospectrophotometry. Use reverse transcriptase-polymerase chain reaction (RT-PCR) to measure the mRNA expression level of acrAB-tolC efflux gene. RESULTS The accumulation of ciprofloxacin in resistant strains was lower than that in susceptible ones, and it could increase to a high level nearly to the susceptible strains. The mRNA level of efflux gene acrA was higher in resistant strains than in susceptible ones. CONCLUSIONS Efflux mechanism is associated with the resistance to fluroquinolones in K. pneumoniae strains isolated in China and CCCP can inhibit its active efflux mechanism , which provides a sensitive method to detect the active efflux system of K. pneumoniae.

Objective Investigate the relation between decrease of apoptosis caused by delayed preconditioning and expression of SMAC and XIAP.Methods Sprage-Dawleyt rats were divided into four groups: control,sham,I/R and IPC/SWOP.The rats in I/R group underwent ischemia for 1 hour by classic artery ligation and reperfusion for 1 hour.The rats in IPC/SWOP group underwent tree cycles of 5-minute ischemia and 5-minute reperfusion 24 hours prior to the index occlusion.Cell apoptosis was measured by flow cytometry,the activity of caspase-3 was also measured.The expression of SMAC and XIAP in cytosol of myocardial cell was measured by Western blot.Results Cell apoptosis rate,activity of caspase-3 and expression of SMAC significantly increased in I/R as compared with control(P

Objective To study the clinical characteristics of multi-drug resistance bacterias(MDROs) isolated from hospitalized patients in the Second People′s Hospital of Longgang District,to provide strategies for the prevention of MDROs infection.Methods The MDROs data of hospitalized patients from January 2015 to December 2016 were analyzed retrospectively.The multi-drug resistance incidence of each bacterias,each types of specimens and each clinical departments were analyzed and compared by SPSS16.0.Results A total of 104 strains of MDROs were isolated,and the top five bacteria were E.coli(32 strains,30.77%),coagulase negative staphylococcus(24 strains,23.08%),pseudomonas aeruginosa(16 strains,15.38%),staphylococcus aureus(10 strains,9.62%),kiebsiella pneumonia(10 strains,9.62%) respectively.There was significant difference in the multi-drug resistance incidence of each bacterias(χ2=20.62,P<0.05),the average incidence was 25.12%,and the top three incidence were E.coli(36.78%),pseudomonas aeruginosa(33.33%),coagulase negative staphylococcus(28.24%) respectively.There was significant difference in the multi-drug resistance positive rate of each types of specimens(χ2=43.68,P<0.05),the average positive rate was 5.84%,and the highest positive rate were wound secretion and pus(11.00%),followed by urine(8.25%).There was significant difference in the multi-drug resistance positive rate of each clinical departments(χ2=40.36,P<0.05),and the highest positive rate were in department of urinary surgery(12.63%),followed by department of gynaecology and obstetrics(11.16%).Conclusion E.coli coagulase negative staphylococcus and pseudomonas aeruginosa were mainly epidemic MDROs,and the MDROs are mainly distributed in urological surgery,obstetrics and gynecolog in this hospital.The occurrence of MDROs should be for the prevention and control strongly in the hospitalized patients with all kinds of trauma or diseases of urinary system and in the clinical department of urinary surgery,gynaecology and obstetrics.

Background:Gemcitabine is the main drug for chemotherapy of advanced pancreatic cancer,however,the prognosis of pancreatic cancer patients has not been changed obviously because of the high innate and acquired resistance of cancer cells to gemcitabine. Aims:To investigate the correlation of DNA repair and expression of human APE1 / Ref-1(apurinic/apyrimidinic endonuclease 1 / redox factor-1),the key enzyme in base excision repair pathway,with the resistance of pancreatic cancer to gemcitabine. Methods:A gemcitabine-resistant human pancreatic cancer cell line SW1990-0. 5 with a resistance index of 9. 32 and its parental cell line SW1990 were treated with gemcitabine. DNA injury was assessed by comet assay. Expressions of APE1 / Ref-1 mRNA and protein were determined by real-time PCR and Western blotting, respectively. Results:In comet assay,after treated with gemcitabine for 24 hours,OTM value of SW1990-0. 5 and SW1990 cells were 0. 32 ± 0. 13 and 26. 96 ± 6. 83,respectively. Expression level of APE1 / Ref-1 mRNA in SW1990-0. 5 cells was 2. 48 ± 0. 49;and expression levels of APE1 / Ref-1 protein in SW1990-0. 5 and SW1990 cells were 1. 57 ± 0. 08 and 0. 84 ± 0. 06,respectively. Statistically significant differences were existed in all these parameters between SW1990-0. 5 and SW1990 cells(P all < 0. 05). Conclusions:DNA repair might be correlated with the resistance of pancreatic cancer to gemcitabine,and up-regulation of APE1 / Ref-1 might contribute to this resistance by its function on DNA repair.