Objective To explore the symptoms of anxiety,family structure,and the distribution features of family about in-patients.And to find out the influence of the family structure on in-patients' symptoms of anxiety.The results would provide evidentiary support and suggestions for in-patients to improve quality of life and for prevention and intervention of anxiety.Methods Family table was used to register the family structure which was classified as the nuclear family,stem/expended family and other classification.The anxiety of in-patients was measured by self-rating anxiety scale.Results A total of 512 cases of in-patients completed the survey,there were 483 valid questionnaires.The rate of effective questionnaire was 94.34％.In family structure of in-patients,core family accounted for 48.65％,stem/expended family was 36.85％.The sex of family structure showed significant difference,the female proportion of the nuclear family was significantly higher than male.Among all in-patients,78.05％ did not have anxiety,low-level,medium-level,and severe anxiety showed no significant differences.The effects of anxiety on family structure had significant differences.The anxiety level of stem family was relatively higher.Conclusions Economic development and the population flow due to industrialization and globalization changed family structure of our country,especially in coastal de-velopment cities with fast economic development,fast-paced and stressful work,and it may affect the influ-ence of Chinese traditional family structure on anxiety of the in-patients.

Objective To investigate the influences on major inflammatory cytokines after co-cul-turing regulatory T cells (Treg) with human umbilical vein endothelial cells ( HUVECs) that were infected with dengue virus type 2 (DENV-2). Methods Peripheral blood mononuclear cells (PBMC) were extrac-ted from concentrated human leukocytes by density gradient centrifugation. Treg cells were sorted by immu-nomagnetic beads. Expression of CD4,CD25 and CD127 molecules on the membrane of Treg cells was detec-ted by flow cytometry to identify the purity of Treg cells. HUVECs pretreated with or without sphingosine-1-phosphate S1P type 1 (S1P1)-specific receptor agonist CYM-5442 for 24 h were first infected with DENV-2 and then co-cultured with Treg cells. Expression of IL-6,IL-8,TNF-α,IL-10 and TGF-β at mRNA level was detected by real-time RT-PCR. Levels of IL-6,IL-8,IL-10 and TGF-β in the culture supernatants were detec-ted by a double-antibody sandwich ELISA. Results The purity of Treg cells was (84. 3±0. 5)%. Expression of NS1 at mRNA level in DENV-2-infected HUVECs first gradually increased and then decreased after reac-hing the peak at 24 h (3. 03±0. 26, P<0. 01). Enhanced expression of IL-6,IL-8 and TNF-α at mRNA level in HUVECs was observed after DENV-2 infection ( P<0. 01). Expression of these cytokines at every time point was decreased after co-culturing DENV-2-infected HUVECs with Treg cells ( P<0. 05),but was still higher than that before infection. CYM-5442 pretreatment decreased the expression of IL-6,IL-8 and TNF-α at mRNA level in DENV-2-infected HUVECs and inhibited the secretion of IL-10 and TGF-β by Treg cells that were co-cultured with DENV-2-infected HUVECs. Conclusion Primary HUVECs infected by DENV-2 can enhance the secretion of IL-10 and TGF-β by Treg cells,and the suppressive cytokines produced by Treg cells can reduce the production of inflammatory cytokines by DENV-2-infected HUVECs.

BACKGROUND AND OBJECTIVEThe p53 as a transcription factor in cell stress was activated to regulate cell cycle and programmed cell death to inhibit tumor growth. Usually, p53 is kept in non-activated state through various mechanisms, including the action of p53 C-terminal negative regulatory sequences. The purpose of the study is to prepare the two types p53 recombinant adenoviruses that carry full-length p53 as well as deletion of negative regulatory sequences at p53 C-terminus and to detect exogenous GFP expression in human lung cancer cell infected-virus by FCM scatter plot.

METHODSUsing pAdEasy-Track vector system the p53 recombinant plasmids was constructed and the homologous recombinants in E. coli was produced. The three kinds of recombinant adenovirus in L293 cells was generated, sequencing proved. Exogenous GFP expression in human lung cancer 801D cells infected-virus was detected by FCM scatter plot.

RESULTSp53 recombinant adenoviruses named Ad-p53(wtp), Ad-p53(del) and Ad-(empty carrier) were produced. Results of sequences indicate that the Ad-p53(del) was deletion of 111 bases before stop codon TGA and of 3 untranslated region at p53, the Ad-p53(wtp) no loss of any p53 base, the Ad-(empty carrier) no p53 sequence. FCM scatter plot indicate the percentage of 801D cells expressed GFP with three kinds of viral infection was almost same and was increased with the virus density. 801D contains ratio of cells with different fluorescence intensity.

CONCLUSIONThe preparation of recombinant adenovirus, Ad-p53(del), pA-p53(wtp) and Ad-(empty carrier). The cells expressed-GFP can be quantitatively detected by FCM scatter plot. It was provide that the reliability of the virus system and accurate method for selecting viruses density to infecting cells.

Adenoviridae ; genetics ; Animals ; Flow Cytometry ; methods ; Genes, p53 ; Green Fluorescent Proteins ; genetics ; Humans ; Mice ; Recombination, Genetic

Objective To investigate the epidemiological information of patients in pre-hospital medical care for our large and medium-sized cities and probe the patients' characteristic. Method The data in 2008 were exported from the computer databases of 8 large and medium-sized cities' emergency medical centers in our country.The thorough records of data were conducted to statistical analysis. Results ( 1 ) The scheduling time, running time, rescue time, returning time, total time and service radius in the pre-hospital medical care group were 2.16± 1.10(min), 14.01 ±6.82(min), 12.12±5.96(min), 14.08± 6.85(min), 42.34± 20.21(min)and 8.50±4.18(km), and the above parameter in the non-death group were 2.19 ± 1.13(min), 14.15 ± 7.14(min),11.60±6.72(min), 14.92 ±6.89(min), 41.86± 19.53(minutes) and 8.63±4.31(Km), and the above parameter in the death group were 2.10± 1.08(min), 13.68 ± 7.14(min), 25.25 ± 12.34(min), 13.75±6.48(min), 54.74 ± 25.47(min) and 7.86± 3.91(Km), and the above parameter in the non-sudden cardiac death group were2.09± 1.03(min), 13.58±6.78(min), 25.53± 12.34(min), 13.60± 6.54(min), 53.79±23.77(min) and 7.67 ± 3.86(Km), and the above parameter in the sudden cardiac death group were 2.12 ±1.02(min), 14.10±7.05(min), 24.79± 12.08(min), 13.79±6.61(min), 54. 80 ± 25. 36( min) and 7.90±3.92(Km) respectively. The scheduling time, running time, returning time and service radius in the death group were less than those of the non-death group, but the rescue time and total time of the former were more than those of the latter respectively ( P ＜ 0.05 or P ＜ 0. 001 ). The scheduling time and returning time didn' t have significant difference between the sudden cardiac death group and the non-sudden cardiac death group respectively ( P ＞ 0.05), but the running time, total time and service radius of the sudden cardiac death group were more than those of the non-sudden cardiac death group, and the rescue time of the former was less than that of the latter respectively ( P ＜ 0.05 or P ＜ 0.001 ). (2)The patients' amount in pre-hospital medical care group, the non-death group, the death group, the non-sudden cardiac death group and the sudden cardiac death group were at most in first quarter, and the least time slice of patients' amount were 4:00～ 6:00, 4:00～6:00, 4:00～ 6:00, 22:00～ 24:00, 2:00～4:00 respectively, and the most time slice of patients' amount were 20:00～ 22:00, 20:00～22:00, 8:00～ 10:00, 2:00 ～ 4:00, 8:00 ～ 10:00 respectively. (3)In 241 876 cases of pre-hospital medical care group, the patients' amount of trauma was at most, whose age grades was by far among21 ～50, and the others in sequence were nervous system, circulatory system, other group, digestive system, respiratory system and poisoning group respectively, whose age grades in nervous system, circulatory system and respiratory system was by far above 51, especially above 70. The patients' age grades in other group and digestive system had two climax age groups, which the one was 21 ～ 30, and the other was above 70. The patients' age grades in poisoning group was by far among 21 ～ 50, which the patients' amount of acute alcoholism was at the most. (4) In 12 568 cases of death group, the death amount of circulatory system, other group, respiratory system, nervous system and digestive system ranked at the lst,2nd,4th,5th 8th respectively, whose age grades was by far above 51, especially above 70,and the patients' amount of sudden cardiac death was at the most in the death amount of circulatory system. The death amount of trauma and poisoning group ranked at the 3rd, 6th respectively, whose age grades was by far among 21 ～ 50. (5)The total amount, the death amount and the sudden cardiac death amount of male patients were more than those of female patients. (6)The percentage of the death group to the pre-hospital medical care group was 5.20%, and the percentage of the sudden cardiac death group to the pre-hospital medical care group was 1.29%,and the percentage of the sudden cardiac death group to the death group was 24.87 %, and the percentage of the sudden cardiac death group to the circulatory system group was 67.33 %. Conclusions ( 1 )The trauma and the sudden cardiac death are the overriding reason of disease and the overriding reason of death in our large and medium-sized cities respectively. (2) It is very important to cut the death rate of the middle-old age patients by strengthening prevention and cure of cardiovascular and cerebrovascular diseases, discerning the critical illness early and improving the level of pre-hospital medical care. (3)It is a strong method to decrease the total amount and the death amount of the trauma, especially in traffic accident, by strengthening safety in production, observing traffic regulation and enhancing the legal awareness.