Purpose To investigate the responsiveness of intracellular oxygen free radical and calcium on acrolein exposure and acrolein-induced cardiomyocytes apoptosis. Methods The viable adult mice cardiac myocytes were isolated by modified langendorff methods. We have examined the intracellular oxygen free radical and calcium concentration using DCF and Fura-2 AM, and the cardiomyocytes viability with WST assay. Are evaluated the DNA ladder pattern and cell apoptotic morphology on the adult mice cardiomyocytes that are exposed to acrolein. Results Our results show that acrolein can increase markedly the intracellular oxygen free radical and calcium concentration, that reach 12 fold and twofold respectively compared to the resting value when the cells were exposed to 1 μmol/L of acrolein. Moreover, the injury induced by acrolein in cardiac myocytes is concentration-dependent. The cardiomyocytes viability treated with 25, 50, 100 μmol/L of acrolein respectively were significantly lower compared to controls (P ＜ 0.01 ). DNA ladder pattern and apoptotic morphological changes were found after being exposed to acrolein in the adult mice cardiomyocytes. Conclusion It is concluded that acrolein induces adult mice cardiomyocytes apoptosis, and it may be due to the increased intracellular oxygen free radicals and calcium concentration.

Objective To investigate the changes in acetylation of histone in the spinal dorsal horn in a rat model of persistent postoperative pain.Methods Ninety-six malc Sprague-Dawley rats,weighing 200-250 g,aged 6-8 weeks,were randomly divided into 2 groups (n=48 each) using a random number table:sham operation group (group S) and persistent postoperative pain group (group PPP).The rat model of persistent postoperative pain evoked by skin/muscle incision and retraction was established according to the method described by Flatters.After the rats were anesthetized with intraperitoneal chloral hydrate,the skin and superficial muscle of the medial thigh were incised and retractors inserted.This tissue was retracted for 1 h.The mechanical paw withdrawal threshold (MWT) was measured at 1 day before operation and 1,3,7,14,and 21 days after operation.Four animals were sacrificed in each group after measurement of MWT at each time point for detection of acetylated histone H3 (Ac-H3) and acetylated histone H4 (Ac-H4) expression (by Western blot analysis) and the number of Ac-H3 and Ac-H4 positive cells in the spinal cord horn (by immunofluorescence histochemistry).Results Compared with group S,the MWT was significantly decreased at 3,7,14 and 21 days after operation,the expression of Ac-H3 and Ac-H4b was significantly down-regulated at 3,7 and 14 days after operation,and the number of Ac-H3 and Ac-H4 positive cells was significantly decreased at 7,14 and 21 days after operation in group PPP (P＜0.05 or 0.01).The MWT,expression of Ac-H3 and Ac-H4b,and the number of Ac-H3 and Ac-H4 positive cells were significantly higher at 21 days after operation than at 14 days after operation in group PPP (P＜0.05).Conclusion Acetylation of histone in the spinal dorsal horn is decreased after operation,which may be involved in the development and maintenance of persistent postoperative pain in rats.

Objective To evaluate the effect of heat treatment on ultraviolet B (UVB)-induced oxidative injury to human melanocytes.Methods Melanocytes were isolated from adult foreskins,and subjected to a primary culture.After 3-4 passages of subculture,the melanocytes were classified into 4 groups:control group incubated at 37 ℃,heat treatment group incubated at 42 ℃ for 1 hour,UVB group exposed to UVB irradiation at 100 mJ/cm2,combination group receiving heat treatment at 42 ℃ for 1 hour followed by UVB irradiation at 100 mJ/cm2.After three successive days of treatment,MTT assay was performed to evaluate cell viability,a biochemical method to determine the activity of superoxide dismutase (SOD) and concentration of malonaldehyde (MDA),and flow cytometry to detect intracellular reactive oxygen species (ROS) and apoptosis in melanocytes.Results The cell survival rate,apoptosis rate,SOD activity,MDA concentration and ROS level were (100 ± 6.14)％,(4.66 ± 0.58)％,(53.39 ± 8.23) U/gprot,(1.09 ± 0.32) mmol/gprot,and 1070.85 ± 42.07 in the control group respectively.UVB exposure induced a significant increase in apoptosis rate (24.14％ ± 2.90％,P ＜ 0.001),MDA concentration (1.65 ± 0.33 mmol/gprot,P ＜ 0.01) and ROS level (1416.45 ± 79.12,P＜ 0.01),but a significant decrease in cell survival rate (50.23％ ± 5.36％,P＜ 0.01)and SOD activity (31.98 ± 1 1.89 U/gprot,P ＜ 0.01) in the UVB group compared with the control group,while the heat pretreatment markedly downregulated the UVB-induced increase in apoptosis rate (14.9％ ± 1.49％,P ＜ 0.001),MDA concentration (1.10 ± 0.26 mmol/gprot) and ROS level (1033.30 ± 68.41,P＜ 0.01),as well as the decrease in cell survival rate (74.12％ ± 6.17％,P＜ 0.01) and SOD activity (51.63 ± 6.55 U/gprot,P＜ 0.01) in the combination group.Conclusion Heat treatment could protect melanocytes from UVB-induced oxidative injury.

OBJECTIVE:To study the incidence of adverse drug reactions(ADRs) and the rational use of Durogesic in in-patients METHODS:37 in-patients treated with Durogesic in Department of Oncology in our hospital were prospectively observed,and its analgesic efficacy and incidence,severity and the outcome of ADRs were evaluated RESULTS:The alleviating effect of Durogesic on carcinomatous pain was 100% Of 37 patients,ADRs were found in 4 cases with a ADR rate of 10 8% CONCLUSION:Durogesic is effective in clinical use,but attention should be paid to monitoring ADRs and avoiding irrational use of drug

Objective To evaluate the effect of multiple adhesion molecules ICAM-1、CD_ 44 V6、CD_ 28 、E-cad and CEA in organic selection of gastric carcinomatous metastasis.Methods Eighty-four gastric carcinoma cases(32 cases without metastases,52 cases with transfer to liver、marrow、lymphonode、lung、kidney or belly cavity),were take operation sample or tissue under gastroscopy,being made cells single by primary culture,then were detected the expression of 5 kinds of adhesion molecules by ABC immunohistochemistry,positive rate was recorded.Results ICAM-1 had high expression in cases with marrow or lung metastasis,while low expression in cases with lymphonode metastasis.CD_ 44 v6 had high expression in cases with lymphnode or liver metastasis.CD_ 28 had high expression in cases with liver、lung or kidney metastasis.E-cad had high expression in cases with marrow metastasis,while low expression in cases with other metastasis.CEA had especially high expression in cases with liver、belly cavity or lymphnode metastasis.Conclusion High or low expression of adhesion molecules ICAM-1、CD_ 44 V6、CD_ 28 、E-cad and CEA plays a certain role in organic selection of gastric carcinomatous metastasis.

Objective To investigate the relationship between miR-411 and breast cancer,and the expression of miR-411 and its effects and mechanism research on proliferation and metastasis in breast cancer.Methods Real-time polymerase chain reaction (RT-PCR) was used to detect the expression of miR411 in breast cancer cells and tissues.Methyl thiazolyl tetrazolium (MTT),clone formation assay and Transwell assay were used to detect the effect of miR-411 expression on the proliferation,migration and invasion in breast cancer cells.The effect of miR-411 on growth factor receptor-bound protein 2 (GRB2) expression in breast cancer cells was detected by RT-PCR and Western blot.The direct effect of miR-411 target on GRB2 was detected by dual luciferase reporter assay.MTT,clone formation assay and Transwell assay detect the effect of GRB2 expression on the proliferation and invasion in breast cancer cells.Detection the effect high expression of miR-411 on GRB2 downstream signaling pathway related molecules expression in breast cancer cell with Western blot.Results The expression of miR-411 in breast cancer tissues was significantly lower than that in adjacent non-cancerous tissues (P ＜ 0.05).The expression of miR-411 in breast cancer cells SK-BR-3,BT-549 and MDA-MB-231 was significantly lower (P ＜ 0.05).Compared to the negative control group,the transfection of miR-411 mimic inhibited the proliferation,migration and invasion of MDA-MB-231 breast cancer cells (P ＜ 0.05).Targetscan showed that miR-411 could bind to GRB2 3'UTR at position 741-747.Compared with the negative control group,GRB2 3'UTR wild-type plasmid and miR-411 co-transfection reduced the fluorescence activity (P ＜ 0.05).Transfection of siGRB2 significantly reduced the expression of GRB2 protein in MDA-MB-231 breast cancer cells (P ＜ 0.05).Compared to the negative control group,the inhibition of GRB2 expression reduced the proliferation and the number of colony formation of MDA-MB-231 breast cancer cells (P ＜ 0.05).Transwell assay showed that transfection of siGRB2 significantly reduced the number of invasive cells in MDA-MB-231 breast cancer cells (P ＜ 0.05).Conclusions miR-411 is related closely to the occurrence and development of breast cancer,miR-411-GRB2-Ras axis is expected to become a new target for biological treatment of breast cancer.

Objective To compare the humoral immune response of BALB/c mice immunized by recombinant plasmids PeDNA3.1-M-NS1 and pcDNA3.1-N-NS1.Methods Dengue type 2 virus(DENV2)NS1 gene were constructed two partial sequences(1-413 bp)of the pcDNA3.1 eukaryotic plasmids and pET28a(+)plasmid for prokaryotic expression,identification,purification and quantification.The BALB/c mice were immunized by pcDNA3.1-M-NS1,pcDNA3.1-N-NS1 recombinant plasmids with adjuvant.Each animal received a primary inoculation and two boosts at 1-week intervals.Then the blood samples of BALB/c mice were collected from different experiment groups at day 7,14,28 and 56,respectively after first immunization.The specific IgM/IgG antibodies for NS1 protein in serum were confirmed by indirect ELISA.And then the activities of the specific protective antibody were determined by cytopathic effect inhibition(CPEI).Results Construction of the pET28a(+)-NS1 m/pET28a(+)-NS1n prokaryotic expression plasmid,SDS-PAGE analysis showed that,NS1 gene partial sequence was expressed,both the relative molecular weight of about 22.3×103:Western blot showed that the protein can bind anti-His tag monoclonal antibody;byNi affinity chromatographywith apurity of 92% protein,on the C6/36 cell toxicity,and can be used ELASA detection.The results showed that the levels of specific IgM/IgG antibody and neutralizing antibody activities were increased in pcDNA3.1-M-NS1 booster immunization group than other groups.The result had been observed longer duration of antibody level in peDNA3.1-M-NS1 booster immunization group.Conclusion Humoral immune response were significantly different between pcDNA3.1-M-NS1 and pcDNA3.1-N-NS1 recombinant plasmid immunized mice groups.

Objective To investigate the requirements of hospitalized patients on humanized nursing in the ward of whole-process humanistic care mode and provide scientific references for continuous improvement of nursing quality.Methods A total of 408 patients from 2 wards of whole-process humanistic care mode.Patients were applied the self-designed questionnaire in combination with interview to understand the patients’ requirements of humanized nursing.Survey was carried out before the inpatients were discharged from the hospital adopting the “Evaluation Table of Patients’ Satisfaction Degree about the Nursing Service” produced and researched by our hospital independently.Results After carrying out the whole-process humanistic care service mode,the inpatients’ overall satisfaction degree and satisfaction rate with humanistic care were respectively 98.2％,97.5％.Conclusions Carrying out the quality care demonstration project and enhancing the concept of humanistic care can promote each other.Carrying out the inpatients’ care needs survey in the ward of whole-process humanistic care mode,integrating humanistic into every aspect of clinical nursing,are the crux of deepening high-quality nursing care as well as a new subject that clinical nurses should confront conscientiously.

for the detection of their virulent abilities simultaneously.

Objective To establish an early,simple and rapid colloidal gold strip method for the detection of IgM against hantavirus nucleocapsid protein in patients with hemorrhagic fever with renal syndrome(HFRS).Methods Purified recombinant Hantavirus nucleoprotein(rNP)was labeled by colloidal gold particles and then sprayed and fixed on fiberglass membrane as the combination pad.Anti-Human IgM(μ-chain specific)antibody produced in goat was fixed in the detection area,and mouse antihantavirus antibody was fixed in the quality control area.Both of them were on nitrocellous membrane strip in tandem.Together with a specimen pad ahead.The conbination pad and the nitrocellous membrane were assembled into a test strip.The colloidal gold strip assay was compared with ELISA for evaluation of specificity and sensitivity.Results The colloidal gold strip tests showed positive in the serum samples from 50 cases of HFRS which was clinically diagnosed and then verified by ELISA within 10-15 minutes.Whereas 30 serum samples of healthy donors have tested negative.Conclusions Our new colloidal gold immunochromatographic test strip method was well concordant with the ELISA assay,but the former was more raoid and simole.It could be used primary medical services.