Objective To investigate the effect of EZH2 knockdown on cell proliferation ,invasion ,migration and apoptosis in hu-man bladder cancer cell line by small interfering RNAs(siRNA) .Methods The siRNA-expressing plasmid targeting EZH2 gene was constructed and transfected into T24 cells .RT-PCR was used to detect the EZH2 gene′s expression at the level of mRNA ;pro-liferation ,invasion and migration of T24 cells were examed in vivo by MTT ,wound healing assay and Transwell chamber migration assay .Finally ,Annexin V-FITC/PI flow cytometric analysis was performed for cell apoptosis .Results The siRNA-expressing plas-mid targeting EZH2 gene successfully inhibited EZH2 gene’s expression in T24 cells .The expression of mRNA was significantly inhibited compared with negative control groups (P<0 .05) .After the transfection of the plasmid 48 hours ,the growth inhibition rate was 37 .9% ,which was higher than the negative control group(P<0 .05) .24 hours after wound healing ,the migration distance of transfected group cell was(1 .37 ± 0 .12) ,which was lower than the negative control group(P<0 .01) .Compared with the nega-tive control group ,invasion capability of EZH2-siRNA group was dropped by 67% (P<0 .01) .48 hours after transfection ,the early and secondary apoptosis rate of T 24 cells were 22 .80% and 3 .60% respectively ,which were higher than the negative contral group (P<0 .01) .Conclusion The siRNA interference EZH2 can significantly inhibit cell proliferation ,invasion and migration of T24 cells ,meanwhile promote its apoptosis .It provides a theoretical basis for further study of bladder cancer gene therapy .

Objective To explore the reliability of patients' sensation of the need to urinate,and to investigate the optimal volume and duration of bladder filling and training method for the stability of bladder filling. Methods From 2014 to 2015,Fifty patients with pelvic tumor were divided into group A and group B,according to whether they had the history of diseases or surgery in the pelvis or urinary system. Both groups received training of the sensation of the need to urinate. The training required patients to drink a fixed volume of water every time they emptied the bladder. The bladder capacity was measured by a bladder scanner ( BladderScan BVI 9400) ,and a rating scale of the sensation of the need to urinate was completed at 30 min,45 min,1 h,and over 1 h when the patient sensed the limit for bladder capacity. The optimal volume and duration of bladder filling or optimal frequency for the training were explored. The paired t-test method was performed for the difference between the predictive value and the measured value,Pearson method was performed for correlation between the sensation of the need to urinate and the measured value. Results In the A and B groups,there was no significant difference between the predicted value and measured value ( A:predicted value and measured value P=0. 777,B:predicted value and measured value P=0. 061) ,suggesting that the measured value could be used to reflect the predictive value. Compared with group B,group A had a higher correlation between the sensation of the need to urinate and the measured value ( rA=0. 812, rB=0. 762).The correlation between the predictive value and the measured value became the highest at 45 min and 1 h ( r=0. 858 and 0. 916) ,and the corresponding bladder filling volume and score of the sensation of the need to urinate were 330-450 ml and 4-6,respectively. The correlation between the predictive value and the measured value increased with the frequency of the training ( r2=0. 914, r3=0. 917, r4=0. 930, r5=0. 951,r6=0. 962) . Conclusions Before radiotherapy,patients with pelvic tumor should received at least 4-6 rounds of the training of bladder filling. Patients should drink 800-1 400 ml of water every time they empty the bladder,and the optimal bladder filling volume and the reliable and stable sensation of the need to urinate will be achieved after 45 min-1 h. For the patients with the history of urinary system diseases or pelvic surgery,the bladder filling volume needs to be measured using a bladder scanner ( BladderScan BVI 9400) during the training before radiotherapy.

Objective To evaluate the application value of the one-stepdiabetes education management system in clinic. Methods The “one-step” diabetes education management system was construct.80 newly diagnosed patients with type 2 diabetes mellitus were selected and randomly divided into the control group and the experimental group with 40 patients in each group.The patients in the experimental group received “one-step”diabetes education management system by diabetes specialist nurses,while patients in the con-trol group received routine diabetic education.The fasting blood glucose (FBG),HbAlc,medical compliance behaviors and other indices were compared between two groups. Results FBG,HbAlc and medical compliance were better,and the risk score of complications was higher in the experimental group than those of the control group,there were significant differences between them. Conclusions Application of “one-step”diabetes education management system in treatment and nursing of patients shows maneuverability and practicality.It has clinical application value and deserves to be spread.

Objective To analyze the association between mtDNA mutations and photodamagc after ultraviolet B (UVB) irradiation. Methods Primary human skin fibroblasts (HSF) and primary human epi- dermal keratinocytes of adult (HEKa) were irradiated by sub-lethal doses of UVB thrice a day for 4-5 days. Thereafter, genomic DNA was extracted from irradiated cells and conventional PCR was applied to detect the frequency rates of 4977 bp and 3895 bp mtDNA deletion. To quantitatively analyze the mutation levels, SYBR Green real-time PCR method was performed. Results In both cell lines, the frequency rates and relative copy number of deletions increased with the cumulative doses of UVB exposure (P<0.05). The prevalence rate of 3895 bp deletion peaked 53.3% and and relative copy number reached (49.63±4.38)×10-5, showing a more intense response to the accumulation of UVB radiation than 4977 bp deletion. In HSF, the minimum cumu- lative dose of UVB radiation was 150 mJ/cm2 for the induction of 3895 bp deletion, and 200 mJ/cm2 for the induction of 4977 bp deletion. It seemed that mtDNA deletion was more readily to be induced by UVB radia- tion in HSF than in HEKa. Conclusions The development and accumulation of mtDNA mutation are intimately related with cumulated UVB dose received by skin cells, and the 3895 bp deletion is more reliable in moni- toring the photodamage caused by UV than 4977 bp deletion. Therefore, the 3895 bp deletion may serve as a biomarker for the detection of photodamagc in skin cells. HSF appear to have an increased susceptibility to UVB radiation, which results in a higher frequency and level of mtDNA mutations compared with HEKa.

Background and purpose:It has been confirmed that homozygous deletion of p16/p15 gene and its co-deletion of p16/p15 genes were related to the occurrence, progress and prognosis of epithelial ovarian cancer. However, the mono-deletion and co-deletion of the genes has been detected with tissue but not in serum DNA of the epithelial ovarian cancer. In this article, we studied the relationship between homozygous deletion of p16/p15 gene and its co-deletion of p16/p15 genes in serum DNA of the epithelial ovarian cancer.Methods:Primers were used to amplify exon 2 of p16 and exon 2 of p15 gene by polymerase chain reaction. Homozygous deletions of the p16, p15 and co-deletion of p16/p15 genes were studied in either serum DNA of 165 patients with epithelial ovarian cancer, their counterpart lymphocytes DNA, serum DNA of 25 benign ovarian cyst or of 15 health donors.Results:The homozygous deletion rates of either p15 or p16 gene were 27.9%(46/165)and 27.3%(45/165)serum DNA in the patients with epithelial ovarian cancer respectively, while the co-deletion rate of p16/p15 genes was 24.2% (40/165). However, the deletions of p15/p16 genes and its co-deletion were not found in serum DNA of the counterpart lymphocytes,25 benign ovarian cyst and 15 health donors (The P values were 0.000、0.000 and 0.000 respectively). The deletions of either p15 or p16 gene for the patients with stage Ⅰ~Ⅱ were 14.3%(5/35) and 11.4%(4/35), 33.3%(25/75) and 32.0%(24/75) for the patients with stage Ⅲ, 29.1%(16/55) and 30.9% (17/55) for stage Ⅳ, respectively. Although there was no significant differences among the groups, the deletion of p15 and p16 genes in the patients with advanced stage were higher than that with early stage. The deletion was not found to be associated with histopathology of epithelial ovarian cancer.Conclusions:Homozygous deletions of the p16, p15 genes and its co-deletion of p15/p16 genes were commonly found in the serum DNA of epithelial ovarian cancer and might be associated with clinical stage of the disease. It was suggested that detection with serum DNA may be used as a micro-invasive approach and the deletion of genes might served as biological markers for the development and prognosis of the patients with epithelial ovarian cancer.

Objective To investigate the photo-toxic effects of indocyanine green (ICG) based on photodynamic therapy (PDT) on vascular endothelial cells in vitro. Methods Human umbilical vein endothelial cells were incubated with ICG with different concentrations in 96-well plate. According to the dose of ICG , they were divided into 6 groups as 0, 50, 100, 150, 200 and 250 μmol/L group. Each group consists of 8 wells, 4 of them were irradiated with a 810 nm laser of total power 24.0 J/cm2. Cell viability was measured by MTT assay 24 h after light irradiation. Morphological changes in cells were observed by light microscope. Results With the irradiation of 810 nm light , there was a significant decrease in relative cell viability in all groups except 0 μmol/L group (all P < 0). With the increase of concentration of ICG, the cell viability dropped rapidly (P < 0). Apoptotic changes can be seen in all groups except 0 μmol/L group and necrosis was the main change in cells of 200 μmol/L and 250 μmol/L group. Conclusion PDT with appropriate concentration of ICG has a strong inhibition to vascular endothelial cells in vitro and ICG-PDT may be a good new therapy for nevascularization.

The methyl group plays an important role in the rational drug design. Introducing methyl into small molecules has become an important strategy of lead compound optimization. The application of methyl in drug design is reviewed in this paper. Methyl can modulate the physicochemical, pharmacodynamic, and pharmacokinetic properties by ortho effect, inductive effect, and conformational effect. It also improves the metabolic stability as a soft metabolic point. In addition, introducing methyl into drug molecules can also be applied as a strategy in new uses of old drugs and generate me-too drugs.

Objective To investigate the clinical liver function protective effect of Shuganning injection for treatment of patients with drug poisoning.Methods One hundred and forty patients with drug poisoning consistent with thecriteria of enrollment into the study were admitted into the Department of Emergency Medicine in the First Hospital of Jilin University-the Eastern Division from January 2015 to August 2016, and they were randomly divided into observation group and control group by the computer generated random numbers, 70 cases in each group. After admission, all the cases were treated with routine treatment including detoxification, removal of toxin, organ protection, symptomatic and supporting treatment, etc. Based on the routine treatment, additionally, Shuganning injection 20 mL+ 10% glucose injection 250 mL, intravenous drip slowly, once a day was given in the observation group; the therapeutic course was 14 days in both groups. The changes of serum glutamic transaminase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBiL) levels were determined before and 7 and 14 days after treatment respectively to evaluate the situation of liver function in the two groups.Results Before treatment, there were no statistical significant differences in the levels of serum ALT, AST and TBil in the two groups (allP > 0.05); with the prolongation of treatment, the above-mentioned indexes of the two groups were gradually reduced, on the 14th day after treatment, they reached the lowest levels, and the degree of decrease in level on the 14th day in the observation group was more significant than that in the control group [ALT (U/L): 32.6±10.8 vs. 98.3±34.5, AST (U/L): 39.4±14.3 vs. 138.5±25.6, TBil (μmol/L): 4.8±1.7 vs. 13.2±2.3, allP < 0.05].Conclusion Shuganning injection has a protective effect on liver function in patients with drug poisoning.

Objective To study the efficacy of RNA interference targeting 5-LOX on 5-LOX and VEGF expression as well as the growth of tumor xenografts in the nude mice,in order to evaluate the value of the clinical application.Methods SW1990 cells were injected into the back of BALB/c nude mice.Once the visible tumors were evidenced about 100mm3,the animals were divided randomly into 4 groups (6 animals/group) and treated with shRNA1 and shRNA2 targeting 5-LOX,negatrve control shRNA (shNC) or control LipofectamineTM 2000 (Lipo) by intratumoral injection.Observing the effect of the shRNA on the growth of tumor xenografts,investigating the expression of 5-LOX and VEGF by immunohistochemistry and RT-PCR.Results Two nude mice were dead in shNC group and Lipo group because of the wasting disease.Other nude mice had no changes in body weight,spirit,appetite,and activity.The growth of tumor xenografts was suppressed potently when administered with shRNA.Compared with shNC and Lipo group,the mean tumor size in groups treated with shRNA was reduced markedly at every point of test time.Between two treat groups,they did not have significant difference.5-LOX and VEGF were both expressed in the pancreatic cancer tissue.The level of the 5-LOX expression in shRNA groups was stronger than that in shNC or Lipo group.The VEGF had the game situation.Between the two treat groups,the difference was not significant.Conclusions RNA interference targeting 5-LOX can inhibit the growth of tumor tumor xenografts in the nude mice by depressing the expression of 5-LOX directly and depressing the expression of VEGF indirectly.

Purpose To evaluate the application value of volume CT perfusion (CTP) protocol with 128-slice CT scanner in patients with unilateral middle cerebral artery (MCA) stenosis or occlusion.Materials and Methods Thirty-five patients with severe unilateral MCA stenosis or occlusion who underwent cerebral volume CTP examination in Wuhan No.1 Hospital between July 2013 and December 2014 were evaluated.The volume CTP parameter maps of cerebral blood flow (CBF),cerebral blood volume (CBV),mean transit time (MTT) and time to peak (TTP) were analyzed.Meanwhile,the dynamic CT angiography (4D-CTA) images were obtained.Results In three patients,CBF,CBV and MTT were observed normal but TTP was delayed.In the other thirty-two patients,CBF in the affected side was lower than that in the contralateral side (but difference without significance:P＞0.05).The increased CBV,prolonged TTP and MTT were also detected in the affected side compared with the contralateral side (difference with significance:P＜0.05).McNemar test showed 4D-CTA and DSA diagnosis of middle cerebral artery occlusion were not significantly different (P＞0.05),and they had better consistency (Kappa=0.861,P＜0.05).Conclusion Brain volume CTP protocol with 128-slice CT scanner provides valuable information about cerebral artery abnormalities and hemodynamic changes in patients with severe stenosis or occlusion of MCA.It has great value for clinical application.