AIM　To quatitatively disclose the relationship between activity and structure of a new class of COX-II inhibitors containing dialkylphenyl-linked heterocyclic moieties. METHODS AND RESULTS Seventeen COX-II inhibitors from literature as a training set were investigated with the aim of developing a 3D-QSAR model using comparative molecular field analysis (CoMFA). To reveal the pharmacophoric pattern, several modes of superimposition were explored. The significant model shows a higher ability to explain and predict the activity of COX-II inhibitors, with the cross-validation RCV2=0.718, non cross-validation R2=0.992, F=260.624, and SEE (standard error of estimate)=0.072. Three compounds were selected as a predicting set, the low deviations of calculated values from the measured ones suggesting a powerful predictive ability of the model. CONCLUSION　The 3D-QSAR explains the dependence of COX-2 inhibition upon the structures of the compounds. Some structure information for design of new COX-II inhibitors with higher activity has been given.

ObjectiveTo investigate the expressions of nerve growth factor (NGF) and its receptors TrkA and p75NTR in dermatofibrosarcoma protuberans and dermatofibroma.MethodsAvidin-biotin immunohistochemical(ABC) method was used to detect the expressions of NGF and its receptors TrkA and p75NTR in paraffin-embedded tissue specimens from 17 cases of DFSP and 15 cases of dermatofibroma.Results NGF and TrkA were highly expressed in both DFSP and dermatofibroma specimens,with no significant difference between the two groups of specimens (x2 =0.11,0.02,respectively,both P ＞ 0.05),while the expression of p75NTR was significantly higher in DFSP than in dermatofibroma specimens(x2 =32,P ＜ 0.01 ).The expression of NGF was positively correlated with that of p75NTR in DFSP(R2 =0.623,P ＜ 0.01 ).ConclusionNGF may play a certain role in the development of DFSP via its high-affinity receptor TrkA and low-affinity receptor p75NTR.

Objective To detect the expressions of nerve growth factor (NGF) and its receptors tyrosine kinase A (TrkA) as well as p75 neurotrophin receptor (p75NTR) in the lesions of lichen planus.Methods Biopsy specimens were collected from the lesions of 32 patients with lichen planus and normal skin of 12 healthy human controls and subjected to paraffin embedding.Immunohistochemical avidin-biotin complex (ABC) method was used to detect the expressions of NGF,TrkA and p75NTR.Results NGF and TrkA,which were located in the cytoplasm of keratinocytes,were strongly or moderately expressed in the lesional skin specimens,but absent or weakly expressed in the normal skin specimens (both P ＜ 0.01).No significant differences were observed in the expression of p75NTR between the lesional and normal skin specimens,or in the expressions of NGF,TrkA or p75NTR among specimens from patients in different age groups,patients of different gender or lesions at different sites (all P ＞ 0.05).There was a positive correlation between the expression of NGF and TrkA in the lesions of lichen planus (R2 =0.535,P ＜ 0.01).Conclusion NGF may play a certain role in the development of lichen planus via its highaffinity receptor TrkA.

Objective To evaluate the impact of the intrauterine device (IUD) insertion on the mental state of women.Methods From Jan.2009 to Jun.2010,a multi-center clinical observational study was performed.Totally 641 women were selected in the six provinces' 18 family planning service stations and hospitals for IUD insertion surgery study.Analysis of the change of women's mental state which was evaluated by symptom checklist-90 (SCL-90) scale before and after IUD insertion surgery.Results Before and after IUD insertion surgery,10 factors' scores in SCL-90 of the observed objects were between 1.1 to 1.2,total scores were 107 ± 27 and 105 ± 25,respectively.Before and after surgery,total average score both were 1.2,the average score of positive items both were 2.1.The difference of the above results were not statistically significance (all P ＞0.05).Preoperative and postoperative,the rate of positive items was 9.2％-19.6％ and 7.7％-17.6％,respectively.In addition to anxiety and fear,the rate of other factors' positive items postoperative was significantly lower than those in the preoperative (all P ＜ 0.05).The incidence of the observed objects postoperative of each factor score,deteriorated was in the range of 4.9％ to 23.0％,improved was in the range of 26.3％-50.1％.The incidence of total scores,deterioration was 28.8％ (166/575),improved was 45.6％ (262/575).The incidence of the average score of positive items,deterioration was 3.7％ (21/575),improved was 52.3％ (301/575).Logistic analysis showed that,in addition to unit level,there were no other significant influencing factors for women' mental state of postoperative (all P ＞ 0.05).Conclusion IUD insertion surgery has no adverse effect on women's mental state.

Objective Through investigating the current situation of scientific research in medicine graduate students, we can provide scientific basis for training postgraduates effectively and reference for graduate education reform. Methods A total of 243 medicine graduate students in Drum Tower Hospital were surveyed and asked to complete the designed questionnaires which included general situation, the objective to study, the situation of participation projects, the management of student, and achievements. Results 33.74％ (82) of students' purpose was confusion, 51.44％ of (125) graduate students engaged in clinical and basic research, the scientific research graduate students and doctoral students participated in the tutor research project more than the professional type graduate students and Postgraduate student (P<0.05). There were significant differences in the number of SCI related papers published by the postgraduates who differ in their ways of training and in different degrees (P<0.05). Conclusion Graduate students are an important part of high-level talents in China. They are the successors of future scientific research. The effectiveness of training postgraduates' scientific research ability will directly affect the future scientific research level of the country. So we should strengthen the cultivation of graduate students' scientific research accomplishment, construct a scientific training model, set up scientific training mode and supervision mech-anism, in order to promote the research level of medical graduate students.

Objective To investigate the inhibitory effects of L. monocytogenes expressing melanomainhibiting activity(MIA)gene on malignant melanoma.Methods The plasmid pERL3-hly-MIA was constructed and used to transform live L.monocytogenes by electroporation.Atier culture.the transformed L.monocytogenes,namely LM-hly-MIA,was harvested for the detection of MIA protein by Westem Blot.A total of 24 mice were divided into three groups,namely control group,LM group and LM-hly-MIA group,to be immunized with physiological saline,L.monocytogenes and LM-hly-MIA,respectively.One day after the immunization.mice were subcutaneously inoculated with 0.1 mL of B16 cells at a concentration of 1×107/mL.One week later.reimmunization was performed.The size and weight of tumors were observed and measured in these mice.Results Enzyme digestion and Western blot confirmed the Successful construction of plasmid pERL3-hly-MIA.The average weight of tumors in mice was 4.33±0.91 gram.3.36 ±0.41 gram and 1.89±0.52 gram,respectively in the control group,LM group and LM-hly-MIA group,respectively(P＜0.05).with the inhibition rates of tumor growth being 22.4%and 61.5%in LM group and LM-hly-MIA group.respectively.Conclusion The attenuated strain of L.monocytogenes expressing MIA gene could evidently inhibit melanoma growth.

Listeria monocytogenes (LM), a Gram-positive facultative intracellular bacterium, can be used as an effective exogenous antigen expression vector in tumor-target therapy. But for successful clinical application, it is necessary to construct attenuated LM stain that is safe yet retains the potency of LM based on the full virulent pathogen. In this study, attenuated LM and recombinants of LM expressing melanoma inhibitory activity (MIA) were constructed successfully. The median lethal dose (LD(50)) and invasion efficiency of attenuated LM strains were detected. The recombinants were utilized for immunotherapy of animal model of B16F10 melanoma. The level of MIA mRNA expression in tumor tissue was detected by using real-time polymerase chain reaction (PCR) with specific sequence, meanwhile the anti-tumor immune response was assayed by flow cytometric analysis and enzyme-linked immunosorbent spot (ELISPOT) assay. The results showed the toxicity and invasiveness of attenuated LM were decreased as compared with LM, and attenuated LM expressing MIA, especially the double-genes attenuated LM recombinant, could significantly induce anti-tumor immune response and inhibit tumor growth. This study implicates attenuated LM may be a safer and more effective vector for immunotherapy of melanoma.

Objective To investigate the inhibitory effect of live-attenuated Listeria monocytogenes (LM)-based vaccines expressing the gene encoding a melanoma differentiation antigen,MART-1,on malignant melanoma and their mechanism.Methods The constructed plasmid pERL3-MART-1 was used to transform live-attenuated LM by electroporation to construct recombinant LM.i.e.△inlB LM-MART-1 and △actA/△inlB LM-MART-1.The half lethal dose (LD50) of attenuated listeria strains was determined by concentration gradient dilution method.C57BL/6 mice were randomly divided into three groups,namely PBS group,△inlB LM-MART-1 group and △actA/△inlB LM-MART-1 group.Mice were inoculated by intraperitoneal injection of O.1 LD50 of each rLM strain or PBS only.One week later,the mice were injected subcutaneously with 1×105 B16F10 cells(a mouse melanoma cell strain)in 200μl of PBS.Reimmunization was performed on day 14 and 21.Subsequently,the growth of tumor and survival of tumor bearing mice were observed.All mice were killed on day 28,and tumor tissue as well as splenocytes were obtained from these mice for the detection of MART-1 gene expression by real-time quantitative PCR and the percentage of CD4+CD25+T cell by flow cytometry.Results The recombinant △inlB LM-MART-1 and △actA/△inlB LM-MART-1 were constructed successfully.The LD50 of △inlB LM and △actA/△inlB LM was lower than LM-EGDe by 100 and 10 000 times respectively.Compared with PBS,the tumor growth was inhibited with △inlB LM-MART-1 by 46.95%(F=6.3,P<0.05),and by 83.96% with △actA/△inlB LM-MART-1(F=37.8,P<0.01).The relative expression level of MART-1 in △inlB LM-MART-1 group and △actA/△inlB LM-MART-1 group was 8.988±0.207 and 11.315±0.445 times that in PBS group (both P<0.05).The percentage of CD4+CD25+T cells in splenocytes was (2.52±0.20)%,(1.14±0.13)% and (0.44±0.15)% in PBS group,△ialB LM-MART-1 group and △actA/△inlB LM-MART-1 group,respectively;the differences were statistically significant between the three groups (all P

Objective:To explore the application of carotid ultrasonography in detecting carotid artery lesions of patients with ischemic cerebrovascular disease(ICVD).Methods:Carotid ultrasonography and vascular reactivity(VR)were performed on 82 patients with ICVD and 40 health controls with Color Doppler Ultrasound.The ultrasonic indexes were statistically analyzed. The intima-media thickness(IMT)and the plaque score,the plaque total area and the cerebral blood flow volume of two groups were compared with each other.Results:Vessel diameter (VD),IMT,peak systolic velocity (PSV),end-diastolic velocity (EDV),mean flow velocity(MFV)in patients with severe stenosis of internal carotid artery and common carotid artery in ICVD group were significantly higher than those in control group,and the differences were statistically significant(P <0.01).VR in patients without stenosis and with mild,moderate and severe stenosis was lower than that in the control group(P <0.05).In ICVD group,IMT,carotid plaque score and plaque total area were significantly higher than those in the control group(P <0.01).Conclusions:The detection on degree of carotid lesions in patients with ICVD via carotid ultrasonography was conducive to understanding the status of ICVD,so as to determine the course and the prognosis.It was significant to clinical treatment of ICVD.

Listeria monocytogenes (LM), a Gram-positive facultative intracellular bacterium, can be used as an effective exogenous antigen expression vector in tumor-target therapy. But for successful clinical application, it is necessary to construct attenuated LM stain that is safe yet retains the potency of LM based on the full virulent pathogen. In this study, attenuated LM and recombinants of LM expressing melanoma inhibitory activity (MIA) were constructed successfully. The median lethal dose (LD(50)) and invasion efficiency of attenuated LM strains were detected. The recombinants were utilized for immunotherapy of animal model of B16F10 melanoma. The level of MIA mRNA expression in tumor tissue was detected by using real-time polymerase chain reaction (PCR) with specific sequence, meanwhile the anti-tumor immune response was assayed by flow cytometric analysis and enzyme-linked immunosorbent spot (ELISPOT) assay. The results showed the toxicity and invasiveness of attenuated LM were decreased as compared with LM, and attenuated LM expressing MIA, especially the double-genes attenuated LM recombinant, could significantly induce anti-tumor immune response and inhibit tumor growth. This study implicates attenuated LM may be a safer and more effective vector for immunotherapy of melanoma.
Animals ; Cancer Vaccines ; genetics ; immunology ; Extracellular Matrix Proteins ; genetics ; immunology ; Listeria monocytogenes ; immunology ; Male ; Melanoma ; genetics ; immunology ; Mice ; Mice, Inbred C57BL ; Vaccines, Attenuated ; genetics ; immunology