Objective To explore effect of DLL4 gene in MCF-7 cells of human breast cancer which was inhibitted by short hair in RNA （shRNA） on inducing apoptosis and chemosensitivity to docetaxel. Methods Specific shRNA was designed in accordance with DLL4 gene and transfected into MCF-7 cells of human breast cancer with liposomal （Lip-shRNA group）， MCF-7 cells transfected with only liposomal as Lip group， and control group without any treatment. Expressions of DLL4 protein in 3 groups were detected by immunohistochemical method， apoptosis and cell cycle distribution were examined by flow cytometry （FCM）. Proliferations and sensitivity of MCF-7 cells to docetaxel in 3 groups were determined by methylthiazoyl-tetrazolium bromide （MTT）. Results The averages optical density value and rate of positive area of Lip-shRNA group were significantly lower than that of other 2 groups （P＜0.01）. The levels of A value at 24h， 48h， and 72h in Lip-shRNA group were significantly lower than that of other 2 groups （P＜0.01）. Rates of cell apoptosis at 24h， 48h， 72h and 96 h in Lip-shRNA group were significantly higher than that of other 2 goups （P＜0.05），and ratio of G2/M was higher too （P＜0.05）. IC50 of Lip-shRNA group to docetaxel was significantly lower than that of other 2 groups （P＜0.05）. Conclusions The RNA interference technology can effectively block the expression of DLL4 gene. Inhibition of DLL4/Notch signaling pathway can lead to proliferation inhibition of cancer cell and a concomitant increase in cells undergoing apoptosis， and can enhance the cell sensitivity to docetaxel. DLL4 may be an important target for therapeutic approach of breast cancer.
Objective To summarize the current advancement of preoperative radiotherapy for rectal cancer. Methods Relevant literatures about current advancement of preoperative radiotherapy for rectal cancer published domesticly and abroad recently were collected and reviewed. Results The lower local recurrence rate and longer disease-free survival time were observed in preoperative radiotherapy， compared with postoperative radiotherapy for rectal cancer. The recurrence rate was higher in short-course radiotherapy， compared with conventionally radiotherapy for stageⅢrectal cancer， but there was no significant difference for stageⅡrectal cancer. The biology molecular such as p53， CEA， Cox-2， EGFR， and VEGF had shown to be radiosensitive. Conclusions The proposal of preoperative radiotherapy for rectal cancer， could be prone to conventionally radiotherapy. There are more screening targets for preoperative radiotherapy in extensive exploration of diverse radiosensitivity. Biology molecular， developed gene expression profiling， and gene chips for rectal cancer may contribute to the individualization treatment.
Objective To investigate the relationship of small airway function with airway sensitivity and reactivity and assess the factors influencingairway hyperresponsiveness (AHR).Methods Data of consecutive subjects with suspected asthma who had a≥20% reduction in FEV1 after ≤12.8 mmol/L cumulative doses of methacholine were analyzed from January 2005 to April 2006.Airway sensitivity was assessed by the cumulative dose of methacholine required to cause 20% reduction in FEV1 (PD20).Airway reactivity was analyzed using the slope of the dose-response curve (DRS).The DRS was defined as the reduction in FEV1 from baseline after the final dose of methacholine inhaled divided by the cumulative dose inhaled.Because of their highly skewed distribution，DRS was logarithmically transformed (log10) for all analysis.Results A total of 184 consecutive subjects aged 16 to 80 years was enrolled.There were 70 male (38.0%) and 114 female (62.0%) subjects.Subjects with higher airway sensitivity，indicated by lower PD20，also had a lower Vmax50% and Vmax25%，and vise versa.PD20 was negatively correlated wit log10DRS (r=-0.874，Plt;0.01).In a simple linear regression model，log10DNS was significantly correlated with FEV1%，Vmax50% or Vmax25% respectively (the determinant r2 were 0.062，0.097 and 0.085，respectively，all Plt;0.01).In a multiple linear regression model that included age，height，and percentage of predicted FEV1，Vmax50% and Vmax25% accounted for 3.9% and 2.6%，respectively，of variability in airway reactivity.The association between Vmax50% and log10DNS was significant in both male and female subjects.The r2 was higher in male subjects.The subjects were divided into three age groups and the association between Vmax50% or Vmax25% and log10DNS was higher in female than in male for age≤25 years，higher in male than in female for 25 -45 years.No association was found for agegt;45 years in both males and females.Conclusions Impaired small airway function is associated with higher airway sensitivity and reactivity to methacholine in subjects with suspected asthma.
Objective To investigate the radiation-sensitizing effects of Ku80 silencing by siRNA interference for A549 lung cancer cells. Methods The sequences of Ku80 siRNA and negative siRNA were chemically synthesized and transfected into A549 lung cancer cells by lipofectamine. RT-PCR and Western bolt analysis were used to determine Ku80 gene expression. The transfected cells in culture dishes were irradiated with X ray at doses of 2 Gy, 4 Gy, 6 Gy, 8 Gy, 10 Gy, respectively. Once all treatments were completed, the cells were processed with the colony formation assay. Results RT-PCR detection showed that Ku80 mRNA levels in A549 lung cancer cells were reduced after transfected with Ku80 siRNA at 24 h, 48 h and 72 h time points. Western blot analysis showed that Ku80 protein content decreased at 48 h and 72 h time points compared with the control group ( P lt; 0. 05 ) . Cloning formation assay indicated that radiosensitivity of A549 lung cancer cells was enhanced after transfected with Ku80 siRNA. Conclusion Ku80 siRNA can effectively inhibit Ku80 gene expression of A549 lung cancer cells, and therefore enhance its radiosensitivity.
Objective To determine if the levels of high-sensitivity C-reactive protein ( hs-CRP)and fibrinogen ( Fbg) can predict the risk of acute exacerbation of chronic obstructive pulmonary disease ( COPD) . Methods hs-CRP was measured by latex-enhanced immunoturbidimetric assay and Fbg was assessed by Von Clauss method. The number of exacerbations was recorded during a 6-month follow-up period. Results Fifty patients with stable COPD were enrolled in the study, of whom48 patients completed the trial and two patients dropped out. During the follow-up, 16 patients had once or more acute exacerbations while other 32 patients had no acute exacerbation. The patients were stratified into two groups ( A-exacerbation, B-no exacerbation) . At the baseline, the patients of the group A had lower FEV1 than thegroup B [ ( 1. 1 ±0. 4) L vs. ( 1. 4 ±0. 5) L, P lt;0. 05] . And the group A had higher hs-CRP and Fbg than the group B [ hs-CRP: ( 4. 6 ±3. 3) mg/L vs. 4. 3 mg/L( IQR 5. 5 mg/L) , P lt;0. 05] ; Fbg: ( 3. 8 ±0. 7) g/L vs. ( 3. 1 ±0. 5) g/L, P lt;0. 05] . Nine of 16 patients with a higher level of hs-CRP( hs-CRP gt;3 mg/L) had acute exacerbations. Seven of other 32 patients with normal hs-CRP level had acute exacerbations. The difference in the acute exacerbations rate between the two groups was significant ( 56. 25% vs. 21. 88% , P lt;0. 05) . All four patients with a higher level of Fbg( Fbg gt;4 g/L) had acute exacerbations. Twelve of 44 patients with normal Fbg level ( Fbg≤4 g/L) had acute exacerbations. The patients with Fbg more than 4 g/L had a higher rate of acute exacerbations( 100% vs. 27. 27%, P lt;0. 05) . After adjusting by age, bodymass index ( BMI) , FEV1 , tobacco consumption and other chronic diseases, the risk of acute exacerbation in individuals with baseline hs-CRP gt;3 mg/L was 9. 33 times higher than those with baseline hs-CRP≤3 mg/L ( 95% CI 1. 870-46. 573) . Conclusion Higher level of hs-CRP is associated with the high risk of exacerbation in patients with COPD.
Objective To investigate the expression and clinical significance of soluble triggering receptor expression on myeloid cells-1（sTREM-1） in sepsis patients.Methods Serum concentrations of sTREM-1，procalcitonin（PCT），tumor necrosis factor alpha（TNF-α） and interleukin-10（IL-10） were measured by enzyme-linked immunosorbent assay，while high sensitivity C-reactive protein （hsCRP） level was detected by immunoturbidimetry in 68 patients with sepsis，40 patients with no-infective SIRS，and 20 normal individuals. The diagnostic and prognostic value of sTREM-1 and its comparison with PCT and hsCRP were analyzed. The sequential organ failure assessment （SOFA） score system was used to evaluate the severity of sepsis. The relationship between sTREM-1, PCT , hsCRP , SOFA score，TNF-α ,and IL-10 of the sepsis patients was analyzed，respectively. Results The differences in the serum concentrations of sTREM-1，PCT，hsCRP，IL-10 and IL-10/TNF-α ratio had statistical significance among three groups（Plt;0.05）. The differences in the serum concentration of TNF-α had no statistical significance among three groups （Pgt;0.05）. However，the serum levels of sTREM-1，PCT and hsCRP in the sepsis group were significantly higher than those in the SIRS group （Plt;0.05）. The receiver operating characteristic curve （ROC） analysis showed the area under the curve （AUC） for sTREM-1，PCT and hsCRP were 0.772 （95%CI 0.674-0.871），0.718 （95%CI 0.601-0.835） and 0.664 （95%CI 0.532-0.797），respectively. The serum levels of sTREM-1 and PCT in the non-survivors were significantly higher than the survivors in the sepsis group （Plt;0.01），but the differences in the serum concentration of hsCRP had no statistical significance between the non-survivors and the survivors in the sepsis group （Pgt;0.05）. There were significantly positive correlations between sTREM-1 and SOFA score，IL-10 or IL-10/TNF-α ratio（r value of 0.453，0.301，0.417，Plt;0.05），but no correlation between sTREM-1 and TNF-α（Pgt;0.05）. There was significantly positive correlation between PCT and SOFA score （r=0.436，Plt;0.05），while no relationship between hsCRP and SOFA score（Pgt;0.05）. Conclusions The serum level of sTREM-1 not only be valuable in the diagnosis of sepsis，but also may be used as a prognostic marker in sepsis，as it can reflect the severity of sepsis in certain degree. Furthermore，sTREM-1 or PCT may be superior to hsCRP in diagnosis，prognostic judgment and severity assessment of sepsis.
Objective To predict clinical chemotherapy sensitivity of primary non-small cell lung cancer（NSCLC） by methylthiazal （MTT） tumor chemosensitivity assay method in vitro and detection of multidrug resistance gene1 （MDR1）, and provide reference for clinical individualized treatment. Methods We selected 80 fresh primary NSCLC samples from NSCLC patients who underwent surgical resection in Zibo Central Hospital Affiliated to Binzhou Medical College between January 2009 and December 2011. There were 46 male patients and 34 female patients with their median age of 54 （29 to 81）years. Viable NSCLC cells obtained from malignant tissue were tested for their sensitivity to cisplatin （DDP）, gemcitabine （GEM）, docetaxe （DOC）, etoposide （VP-16） ,and vinorelbine （NVB） using MTT assay in vitro. Fluorescent quantitative reverse transcriptase-polymerase chain reaction （RT-PCR） was used to analysis the expression level of multidrug resistance gene1 （MDR1）. Results After exposure to antitumor drugs, morphologic changes, decrease of metabolic activity, and apoptosis were detected in NSCLC cells. MTT results showed that different individual cancer cells had different chemosensitivity to antitumor drugs, and cancer cells also had different chemosensitivity to different antitumor drugs. Inhibitory rates of cancer cells exposed to DOC, GEM, and VP-16 were significantly higher than those of cancer cells exposed to DDP and NVB （42.5%±9.5%, 40.5%±6.5%, 38.4%±7.6% versus 31.5%±8.5%，32.5%±7.8%, P＜0.05）.The positive rate of MDR1 in tumor tissues was 40.0% （32/80）. The expression of MDR1 was not associated with tumor histological type, degree of differentiation, lymph node metastasis and TNM stage. The expression of MDR1 was associated with resistance to NVB （χ2=5.209，P=0.022），GEM （χ2=4.769，P=0.029），VP-16 （χ2=4.596，P=0.032），and DDP（χ2=6.086，P=0.014）, but not associated with resistance to DOC（χ2=0.430，P=0.512）. Conclusion MTT chemosensitivity assay can effectively predict clinical chemotherapy sensitivity. Detection of MDR1, together with MTT chemosensitivity assay, can more accurately predict NSCLC chemosensitivity and be a guide for individualized chemotherapy of NSCLC.
【Abstract】 Objective To increase the viscosity of chitosan/glycerol phosphate（C/GP）and to improve its preparation technique in order to develop the appl ication range of C/GP. Methods Chitosan was treated by high-pressure vapor steril ization in order to prepare high viscous C/GP（HV-C/GP）and prepare C/GP by standard methods. The rheologic changes of HV-C/GP and C/GP were detected dynamically by the Gemini rheometer. The initial solution viscosity, gelation temperature and gelation time were evaluated after the viscosity of the materials were increased. Two gelation materials were placed into continuous flow thermostated cells under the same condition and harvest them at predetermined time intervals, 1st, 2nd, 5th, 10th and 25th days, then they were dried, weighed and the mass loss rate was calculated. Ultrastructure of the freeze-dried samples was visual ized by the scanning electron microscope. Results The initial viscosity of C/GP was 1.81 Pas and that of HV-C/GP was 17.24 Pas. The latter one increased 10 times as well as the former one. The gelation temperature of C/GP was 37°C and that of HV-C/GP was 34°C. There was no remarkable difference in gelation time between them. The mass loss rate of HV-C/GP at first day was 72.5% and at 25th days was 90.8%, while that of C/GP was 55.4% and 78.2%. Porous network structure was observed by the scanning electron microscope in both of them. The pore diameter of C/GP was 50-100 μm and that of HV-C/GP was 30-50 μm, which was obviously smaller than the former. Conclusion The viscosity of HV-C/GP prepared by improved technique obviously increases and the thermosensitivity has no significant changes. The degradation time of HV-C/GP in vitro lengthens. The micrographs show that the HV-C/GP gels are porous and the pore diameter are smaller than C/GP.
Objective To analyze the etiology trends, etiological characteristics treatment effects of endophthalmitis in 10 years which from 2000 to 2009 in our hospital. Methods 165 patients (172 eyes) of endophthalmitis in hospital from January 2000 to December 2009 were enrolled in this study. The patients included 122 males and 43 females. The median age was (39.0plusmn;23.7) years. The best visual acuity (BCVA) was non light perception in 16 eyes, light perception 0.05 in 135 eyes, ＞0.05 in 12 eyes. Nine children did not have visual acuity records. There were 141 patients (85.45%) with exogenous endophthalmitis which including 89 patients of trauma, 43 patients after intraocular surgery and nine others; 24 patients (14.55%) with endogenous endophthalmitis. 113 eyes were received intravitreal injection with vancomycin 0.1 ml (10 mg/ml). 152 eyes had pathogenic microorganisms culture records of aqueous humor or aqueous humor and vitreous. The positive records were given drug sensitivity test. The types of endophthalmitis, pathogenic microbial culture and drug sensitivity test results and treatment effects were analyzed. Software of SPSS was used for the clinical data statistical analysis in this study. Results Of 152 eyes with a record of aqueous/vitreous samples pathogenic bacteria culture, 42 eyes (27.63%) had a positive result. In which, 28 eyes showed positive in bacteria culture, 12 eyes was positive in fungus culture and two eyes had a positive in culture of fungus and bacteria growing. The culture positive rate was higher in exogenous endophthalmitis than that in endogenous endophthalmitis (chi;2=4.721 9,P=0.029 8).Most of the G+ positive bacteria were resistance to cephalosporin and quinolones except levofloxacin; but sensitive to vancomycin, rifampin and sulfamethoxazole. The intervention effect was more available for postoperative endophthalmitis than that for traumatic endophthalmitis and endogenous endophthalmitis, the difference was statistically significant（chi;2=38.941 3，P=0.000 0).The BCVA of 23 cases was ＞0.05 after the treatment, compared with before the treatment, the difference was statistically significant (chi;2=3.867 3,P=0.049 2).Compared the ratio of past five yearsprime; to that of recent five years, endogenous endophthalmitis was increased from 7.89% to 20.23% (chi;2=5.014 0，P=0.025 1); postoperative endophthalmitis decreased from 27.63% to 24.72%, and traumatic endophthalmitis decreased from 60.53% to 48.31%, other causes linked endophthalmitis raised from 3.95% to 6.74%. Conclusions In recent 10 years (from 2000 to 2009), the patients with endogenous endophthalmitis are growing. The positive rate of pathogenic agent culture is low, but the culture positive rate of the specimens from endogenous endophthalmitis is higher than that from exogenous endophthalmitis.The treatment was more available for postoperative endophthalmitis than that for other two types of endophthalmitis. The general visual prognosis is poor.
Objective To determine the correlation between central macular thickness (CMT) and the visual function in patients iwht macular edema (ME). Methods The clinical data of 42 eyes of 40 patients with ME which were examined by optical coherence tomography (OCT) and microperimetry (MP-1) were retrospectively analyzed. In 40 patients (42 eyes), diabetic ME (DME) was in 27 eyes，branch retinal vein occlusion was in 11eyes, and central retinal vein occlusion was in 4 eyes. All of the eyes had undergone OCT，MP-1 and best-corrected visual acuity (BCVA) test. Central macular thickness (CMT) was measured by fast macular scans using OCT. Retinal sensitivity (MS) and fixation patterns were evaluated by Mp-1. The position was chosen :2 disc diameters (DD) temporal to the disc and one third of a DD inferior to the centre of the disc. Results The correlation between CMT and BCVA is not significant (r＝－0.429，P=0.069) as well as the correlation between CMT and MS (r＝－0.433，P=0.058). The difference of CMT between the unstable and stable group was significant (F=3.262, P=0.039）. The difference of CMT between the central fixation group and preferred retinal locus (PRL) group was significant (F=3.173, P=0.044). Conclusions BCVA and MS have no significant correlation with CMT. When CMT increases, the fixation stability decreases, fixation location.changes, and PRL occurs.