Aim To explore antitumor mechanism of a recombinant vaccinia virus containing the human CEA-cDNA (rV-CEA). Methods C57/BL mice were immunized three times with rV-CEA. Six weeks later, the macrophages(MΦ s)and splenocytes from rV-CEA-immunized donors were transferred to CEA＋ -HePa tnmor-bearing recipients,Meanwhile, the antitumor effects of these donor's MΦ s and splenocytes and that of the recipient's splenocytes were detected in vitro. Results The MΦ s and splenocytes from rV-CEA-immunized donors possessed strong antitumor activity in CEA-positive tumor-bearing recipients. The in vitro antitumor effect of splenocytes from mice inoculated with MΦ s from rV-CEA-immunized donors were markedly stronger than those from W-VV-immunized donors. However,the in vitro antitumor effect of the MΦ s from rV-CEA-immunized donors was the same as those from W-VV-immunized donors. Conclusion It is demonstrated that antitumor activity induced with rV-CEA may be mediated mainly by antigen present cells (the MΦ s), which activated tumor-specific T cells to kill tumor cells.

Objective To examine the effects of Shenqi Bufei Tang decoction on the expression of histone deacetylase-2 ( HDAC2) and nuclear factor-κB p65 ( NF-κB p65) in the airway smooth muscle tissues of COPD rats with lung-qi deficiency syndrome. Methods A total of 40 male rats were randomly divided into normal control group,model control group,Shenqi Bufei Tang decoction group,and aminophylline group.The COPD rat model with lung-qi deficiency syndrome was established by intra-tracheal instillation of lipopolysaccharide (LPS) and passive smoking for 28 days.Pathological changes of lung tissues were ob-served under the light microscope and the thickness of the small airway wall and airway smooth muscle ( ASM) layer analyzed by the image analysis.Immunohistochemistry,real-time PCR and Western blotting were used to detect the expressions of NF-κB p65 and HDAC2 in ASM. Results The thickness of the airway wall and ASM,and the expression levels of NF-κB p65 mRNA and protein were significantly increased in the model control group when compared with those in the normal control group ( P<0.05) . They were significantly reduced and the expression levels of HDAC2 mRNA and protein were markedly increased in Shenqi Bufei Tang decoction group and aminophylline group,which were compared with those in the model group (P<0.05).There were no sig-nificant differences in these indices between Shenqi Bufei Tang decoction group and aminophylline group (P>0.05). Conclu-sion Shenqi Bufei Tang decoction can inhibit the proliferation of ASM in COPD rats with lung-qi deficiency syndrome,which may be associated with the increased expression of HDAC2 and decreased expression of NF-κB p65.

OBJECTIVEThe effect of triptolide on the DNA methylation level of MMP-9 gene and the mRNA expression of tissue inhibitors of met-alloproteinases (TIMPs) were examined in human fibrosarcoma HT-1080 cells to explore the molecular mechanisms involved in the anticancer activity of triptolide.

METHODHT-1080 cells were cultured in MEM containing 10% newborn calf serum and 1% penicillin-streptomycin. Triptolide was dissolved in dimethyl sulfoxide (DMSO) at a concentration of 1 goL-1 and stored at -20 degrees C. Triptolide was freshly diluted with culture medium perior to use and directly added to cell cultures at the indicated concentration, and incubated for 72 hours at 37 degrees C in a humidified atmosphere with 5% CO2, with changes of reagents every 24 hours. Methylation specific PCR (MSP)was applied to assess the methylation status of MMP-9 gene promoter, and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was employed to measure the mRNA expression of tissue inhibitors of metalloproteinases (TIMPs) in human fibrosarcoma HT-1080 cells after 72 hours of treatment with 6 nmol x L(-1), 12 nmol x L(-1) or 18 nmol x L(-1) triptolide, respectively.

RESULTSThe methylation index of MMP-9 gene promoter was statistically elevated in HT-1080 cells after 72 hours of treatment with 18 nmol L(-1) triptolide, compared with those in controls (0.61 +/- 0.10 vs 0.39 +/- 0.10, P < 0.05), while no significant difference was noted between 6 nmol x L(-1) or 12 nmol x L(-1) triptolide treated HT-1080 cells and controls (0.40 +/- 0.15 vs 0.39 +/- 0.10, 0.46 +/- 0.20 vs 0.39 +/- 0.10, respectively, both P > 0.05). The mRNA expression of TIMP-1, -2, -3 or -4 was not significantly changed in HT-1080 cells after 72 hours of treatment with the indicated concentrations of triptolide, respectively compared with those in controls (all P > 0.05).

CONCLUSIONThe results demonstrated that triptolide upregulates the methylation level of MMP-9 gene in HT-1080 cells in vitro.

Antineoplastic Agents, Alkylating ; pharmacology ; Cell Line, Tumor ; DNA Methylation ; drug effects ; Diterpenes ; pharmacology ; Epoxy Compounds ; pharmacology ; Fibrosarcoma ; drug therapy ; genetics ; metabolism ; Gene Expression Regulation, Enzymologic ; drug effects ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Phenanthrenes ; pharmacology ; Tissue Inhibitor of Metalloproteinases ; genetics ; metabolism

OBJECTIVE:To provide reference for promoting smoothly implementation of deprescribing. METHODS:A total of 335 elderly patients with chronic disease were randomly selected from 4 communities and 9 village clinic of a Chongqing community health service center. Questionnaire survey was conducted about general information of respondents,disease and drug use condition and attitude to deprescribing by using Australian Patients'Attitudes Towards Deprescribing questionnaire as reference. The results of questionnaire survey were analyzed statistically. The cognitive function was assessed by Hastgawa Dementia Scale. The degree of weakness was evaluated by Chinese edition of Edmonton Frail Scale. The quality of life was evaluated by Chinese edition of European Five Dimensional Health (EQ-5D) Scale. RESULTS:Totally 311 valid questionnaires were obtained,with effective recovery rate of 92.8％. Overall,311 participants were recruited,with a median age of 70 years,5 types of median disease (hypertension was most common) and 5 types of median medication (mainly calcium channel blocker and non-insulin hypoglycemic agent);54.7％(170 cases)participants took multiple drugs. The attitudes towards deprescribing were that 39.5％ of respondents believed that they took too many drugs;73.9％ had a desire to reduce their drugs;83.6％ reported that they would be willing to reduce drugs if their doctor said it was possible. The patients taking multiple drugs preferred to use less drugs (P=0.001) and reduce current drugs (P=0.001),and were more willing to reduce drug cost by reducing drug use;they were also more worried about drug side effects than those without taking multiple drugs (P=0.005). CONCLUSIONS:The phenomenon of elderly chronic diseases patients with multiple diseases and multiple drug use are popular in community. It is necessary to simpilfy prescription. Most elderly patients would like to reduce their medications if doctors say it is possible;the patients with multiple drug use are more willing to simplify prescription than those without multiple drug use.

Objective To investigate the methods and internal quality control ( IQC ) leucorrhea routine examinationin clinical laboratories of medical institutions in Guizhou Province.Methods In 2009, 97 clinical laboratories were randomly selected for the first investigation.At the same time, staffs in theinvestigated laboratories were educated on the importance of IQC.The second investigation of the same items was carried out in 2011 inthe same laboratories.The results of the two investigations were analyzed byChi-square test.Results 2009 and 2011 numbers of laboratories thoseonly used normal saline suspension method for leucorrhea examination were 17and 16 (χ2 =0.037, P >0.05 ) respectively, used bothnormal saline and 10%KOH suspension methodswere 16and 2(χ2 =12.003,P<0.01), used staining method were 64and 79(χ2 =5.488,P<0.05), both used suspension and staining methods were 60and 73(χ2 =4.041, P<0.05), used normal salinesuspension method combined with Wright stain and Gram staining methods were3and 28(χ2 =23.996,P<0.01) respectively.Numbers of Laboratoriespracticing IQC were 2and 88in 2009 and 2011 respectivly(χ2 =153.293,P <0.01).Conclusions Currently, the most common used method for leucorrhea routine examination is suspension.Through the investigations and education, the quality ofleucorrhea routine examination was improved in Guizhou Province.

Objective: To explore the effect of the umbilical cord mesenchymal stem cells(UC-MSCs) on the pulmonary fibrosis in silicosis rats. Methods: SPF male Sprague Dawley rats were randomly divided into control group, silica model group and UC-MSCs treatment group with 12 rats each group. SiO₂ intra-tracheal injection(0.5 ml of 50 mg/ml/rat) were applied to silica model group and UC-MSCs treatment groups. After that UC-MSCs treatment group received 1 ml UC-MSCs suspension (3×10⁶ cells/ml) by tail vein injection on the 29th, 36th, 43th and 50th day after exposure to the first silica suspension. On the 60th and 75th day after exposure to silica suspension, all animals were examed for pulmonary CT. Then the rats were euthanized on 75th day after the first exposure to silica.Lung's histopathological examination of the rats from all the groups were carried out. The content of hydroxyproline in lungs, TGF-β1 and IL-6 in serum were examined. Results: The lung's histopathological examination showed no obvious inflammatory cell and no fibrosis in the lung tissue of the control group, there were a lot of inflammatory cell aggregation and collagen fiber deposition in silica model group, while in the UC-MSCs intervention group and treatment group, there were less inflammatory cells and collagen fiber. The rats from silica model groups had higher HYP, TGF-β1 and IL-6 than the rats from UC-MSCs treatment group and control group. Lung fields of rats in the control group were clear and no obvious high-density shadow. Different-sized granular high-density shadows or reticular fibrous shadows were found diffusely distributed in the lungs of the rats in silica model group. Lung field of rats in UC-MSCs intervention group and treatment group were less high density shadows, and more clear. Conclusion UC-MSCs can alleviate the pulmonary fibrosis in silica model rats through regulating the secretion of some fibrosis related cytokines.

Objective: To explore the international development trends and research hotspots of outdoor activities affecting the progression of children’s myopia, and to provide a reference for researching on effective ways to prevent children’s myopia. Methods: Totally 291 relevant documents included in the "Web of Science" core set database were used as research objects, and CiteSpace software was used for visual analysis. Results: At present, the publications in this field were mainly in the United States(81), China(80), Australia(76), and Singapore(33); the top three research institutions were "Natl Univ Singapore"(29), "Australian Natl Univ"(27), "Capital Med Univ"(25); the main authors were "Saw SM", "Morgan IG", "Mitchell P". The field has been developed on the basis of "Ophthalmology", "Public, Environmental and Occupational Health", and has been integrated into 32 disciplines. The research content included "exploration of high risk factors for the progression of children’s myopia" and "outdoor activities", "intervention in children’s progression of myopia" and "longitudinal tracking of children’s vision development". Randomized clinical trials that longitudinally track the correlation between changes in eyeballs and the progression of myopia and the effects of outdoor activities on the biological characteristics of children’s eyeballs have become a hot topic in this field. Conclusion Research on the effects of outdoor activities on the progression of myopia in children has increased dramatically. The study of increasing outdoor activities to interfere with the progression of myopia in children and the vertical tracking of key factors affecting the biological characteristics of children’s eyeballs have become the current international trends.

Objective:To study the role of ultraviolet fluorescence detection technology in personal protective equipment education (PPE) and training.Methods:A study was designed to inspect the risk of self-contamination during PPE doffing between 77 healthcare workers. Used a fluorescent tracer slurry which put on the hands, chest, abdomen, knees to simulate the contaminations. Self-contamination of scrubs and skin was measured using ultraviolet light visualization respectively.Results:According to the uv-fluorescer simulating study, 43 (55.8%) of the medical staff had contamination after the removal of PPE, and the main sites of contamination included: left side of the abdomen 11 (11.70%), left side of the chest 9 (9.57%), left forearm 6 (6.38%), left foot instep 6 (6.38%), neck 6 (6.38%), right shoulder 5 (5.32%), etc. Among them, the frequency of simulated fluorescence pollution in the group with working years less than 6 years was less than that in the other groups, and the difference was statistically significant compared with the group of 11-15 years ( t value was -3.685, P value was 0.001 ). Conclusion:Ultraviolet fluorescence labeling detection technology can directly, quickly and effectively evaluate and feedback the key contaminated parts in the process of using PPE, which can provide detailed evidence for redesigning PPE and improve the PPE training process to reduce the contamination.

Objective To establish a mouse embryonic stem cell test (mEST) model and human embryonic stem cell test (hEST) model, to evaluate the embryotoxicity of di(2-ethylhexyl) phthalate (DEHP). Methods We developed mEST and hEST models according to the European Centre for the Validation of Alternative Methods (ECVAM). We used penicillin G (PN-G) as the standard negative reference and 5-fluorouracil (5-FU) as the standard positive reference, respectively, to verify validity of the models. Based on model validity, mouse embryonic stem cells D3 (mESC-D3), mouse Balb/c-3T3 (3T3), and human embryonic stem cells H9 (hESC-H9) were administered different concentrations of DEHP (15.6, 31.2, 62.5, 125.0, 250.0, 500.0, and 1 000.0 μg/ml) for 7 days. A cell counting Kit-8 was used to detect the 50%inhibitory proliferation concentration (IC50) of mESC-D3 cells, 3T3 cells, and hESC-H9 with DEHP. mESC-D3 and hESC-H9 were treated with DEHP (15.6, 31.2, 62.5, 125.0, 250.0μg/ml, and 500.0μg/ml) for 10 days based on the cytotoxicity results. At day 10, the expression of cardiomyocyte differentiation gene alpha-myosin heavy chain (α-MHC) was detected by real-time PCR and the 50% inhibition of cardiomyocycte differentiation (ID50) determined. Based on the values of IC50 and ID50, functionsⅠ,ⅡandⅡcould be calculated by three linear discriminant functions in the EST model and the embryotoxicity of DEHP described by comparing the three functions. Results Nontrophoblast lineage both ES cells were cultured under optimal conditions and highly expressed hESC markers OCT4 , SSEA4, and TRA-1-60. The embryoid bodies formed were uniform in size and shape, and these results were highly repeatable. The PN-G and 5-FU results coincided with the prediction by ECVAM. Validation of our EST models was satisfactory. Results of the three endpoints of DEHP in mEST were 197.3 μg/ml (IC50 3T3), 210.0 μg/ml (IC50 D3) and 246.8μg/ml (ID50 D3). DEHP was evaluated to be a nonembryotoxic compound based on values of functionⅠ(7.78), functionⅡ(7.58) and functionⅢ(-7.79). The three endpoints of DEHP in hEST were 195.4μg/ml (IC50 3T3), 184.8 μg/ml (IC50 D3), and 84.3 μg/ml (ID50). By comparing the values of function Ⅰ (3.21), function Ⅱ (5.77), and function Ⅲ (-6.46), DEHP was evaluated to be weakly embryotoxic. Conclusion DEHP was determined to be a nonembryotoxic compound by mEST and weakly embryotoxic by hEST. Therefore, hEST is a more sensible model for the evaluation of DEHP embryotoxicity.

Objective:To explore the difference of methylation of circRNA related m6A in early inflammation of silicosis and to elucidate the underlying molecular mechanism of circRNA involved in the process of silicosis.Methods:The activation markers of macrophages were detected by Western blotting (WB) in THP-1-derived macrophages. The cell viability was detected with CCK8, by which the stimulation concentration and time of silica were determined. The methylation of total RNA was determined by colorimetry, and the expression of RNA m6A methylase, demethylase and reading protein were detected by Western blotting in mouse model of silicosis. The differential expression of m6A modified circRNA in lung tissues form silicosis and control mice was obtained through Arraystar m6A circRNA epigenetic transcriptome Chip and verified by RT-PCR.Results:The concentration of SiO 2 at 50 μg/cm 2 had the most significant effect on the activation markers and activity of macrophages. Compared with the control group, SiO 2 increased the total RNA m6A level of macrophages, and there were significant differences in the expression of methylase METTL3 and reading protein YTDHF3. High throughput sequencing analysis showed that compared with the control group, the methylation levels of 132 circRNA m6A in the lung of silicosis model mice were increased, while the methylation levels of 296 circRNA m6A were decreased, and then the target circSLC2A13 was screened based on the basic expression. Further verification showed that SiO 2 significantly increased the expression of circSLC2A13 and m6A modification in macrophages. Conclusion:The methylation of circRNA m6A is involved in the activation of macrophages in early inflammation of silicosis.