Objective To investigate the functions of Monocyte chemotactic protein-induced protein 1 (MCPIP1) in human breast cancer cell line MDA-MB-231.Methods MDA-MB-231 cells were transfected with GFP-tagged MCPIP1 by Tet-on inducing expression system.Endogenous MCPIP1 was knocked down by stable expressing shRNA.MTT assay was performed to measure the growth of MDA-MB-231 cells after overexpression or knockdown of MCPIP1.FACS method was used to analyze cell cycle in MDA-MB-231 cells.Real-time PCR was used to test the expression of cell cycle-related mRNAs expression and their half-lives.RNA-IP experiment was conducted to detect the mRNA directly enriched by MCPIP1.Luciferase assay was performed to determine whether the mRNA decay was mediated through 3′UTR.Results MCPIP1 overexpression significantly inhibited cell proliferation(P<0.05), while knockdown MCPIP1 promoted cell proliferation with statistical significances (P<0.05).MCPIP1 induced cell cycle arrest in MDA-MB-231 with statistical significance (P<0.01).MCPIP1 overexpression reduced the half-lives of cell cycle mRNAs (CDK2,CDK6,cyclin D1,cyclin E1,respectively) with significance (P<0.01).In addition, cell cycle-related mRNAs were able to be pulled down by GFP-MCPIP1 but not isotype IgG(P<0.05).Compared with control vector, MCPIP1 significant suppressed luciferase activities of all four 3′UTR reporters (P<0.05).Conclusions MCPIP1 functions as a tumor suppressor in human breast cancer cell line MDA-MB-231 through inducing G1 cell cycle arrest.

Objective To explore the effects of simvastatin in the treatment of hyperlipidemia fatty liver. Methods Animal models were set up by feeding high caloric diet. Liver index (liver weight/body weight), liver function, blood lipid, liver lipid, the content of MDA in serum and liver were assayed before and after therapy with simvastatin. Results Liver index, blood lipid, liver lipid, and the content of MDA in liver as well as in serum increased significantly ( P

Objective To explore the impact of transtheoretical model on sun protection behaviors of soldiers in plateau area.Methods A total of 208 male soldiers trained in 4 200 meters altitude and medically supported by our hospital were randomly divided into the experimental group(n=104) and the control group(n=104).The experimental group received health education based on the transtheoretical model,and the control group received routine health education.Score of sun protection behaviors questionnaire,sunscreen using behavior and score of the Dermatology Life Quality Index (DLQI) before and after the intervention between two groups were compared.Results Score of sun protection behaviors questionnaire in the experimental group was higher than that in the control group (P＜0.05),sunscreen using behavior in the experimental group was better than the control group(P＜0.05),score of the Dermatology Life Quality Index in the experimental group was lower than the control group(P＜0.05).Conclusion Health education guided by the transtheoretical model can improve the plateau soldiers' sun protection behaviors,and it is an effective method of health education.

Objective To explore the method of primary isolation, cultivation and identification of rat brain microvessel pericytes. Methods The brain tissue of 10 3 week-old Wistar rats was separated sterilely. The brain microvessel fragments were separated using two-step enzyme digestion and one-step gradient centrifugation and were seeded in 35-mm dishes for primary culture. The cell morphology was observed by phase contrast microscopy; the immunofluorescence assay was used to identify the associated antigns, such as the α-smooth muscle actin (α-SMA), neuron-glial antigen 2 (NG2), von Willebrand factor (vWF), and glial fibrillary acidic protein (GFAP). Methyl thiazolyl tetrazolium was used to determine the cell growth curve. Results Pericytes climbed out from the adherent brain microvascular fragments around,showing polygonal, and the cell fusion was 95％ after 12-14 days. Immunofluorescence staining revealed that the molecular markers of the pericytes α-SMA and NG2 related antigens showed double positive, while the vWF and GFAP related antigens showed double negative and the cultured cells were confirmed as brain microvascular pericytes. The growth rate of primary cells was slower. The passage cells entered into logarithmic growth phase after 36 to 60 hours and entered into plateau phase after 72 to 108 hours. Conclusions This method may successfully isolate rat brain microvascular pericytes with higher purity.

Aryldialkylphosphatase ; physiology ; Humans ; Organophosphate Poisoning ; enzymology

Objective To explore the perioperative nursing of patients from high altitude earthquake-stricken regions. Method The perioperative nursing histories of 18 patients from high altitude earthquake-stricken regions were retrospectively analyzed to summarize the nursing experience.Results No wound infection,crush injury or complications occurred.Sixteen patients had good functional recovery,1 patient was at the stage of recovery about limb movement and another one was also at the recovery period from lumbar vertebral burst fracture complicated with nerve injury.Conclusion Close observation,psychological nursing and functional exercise are important for improved success rate of treatment for the patients from the earthquake-stricken regions.

Objective To study the application effect of health education guided by the theory of cross-theoretical model in the training of the first aid training in the basic units of the plateau. Methods Choosing 386 long-term presence of plateau soldiers, and divided them into the control group and the experimental group, the control group was educated by traditional method, in the experimental group, the health education was carried out with the trans-theoretical model, nursing experts gave them targeted intervention. The number of groups at each stage was analyzed in different stages, the same to examination results and pass rate. Results After 6 months of intervention, there was a significant improvement in the mastering of the battlefield Five emergency technology of the experimental group and the scores were better than those of the control group (P<0.05). Theoretical results showed that pass rate of the control group was 89.1%(172/193), the experimental group was 99.5%(192/193), the difference between the two groups was statistically significant (χ2=7.129, P=0.039). In the survey of technical operation, ventilation, hemostasis, bandaging, fixing and handling examination pass rate of the control group respectively were 88.1%(170/193), 89.1%(172/193), 89.1%(172/193), 88.1%(170/193), 88.6%(171/193), the experimental group were 99.5%(192/193), 97.9%(189/193), 99.5%(192/193), 96.9%(187/193), 97.4%(188/193), there were significant differences between the two groups (χ2=7.896-12.759, P=0.001-0.043). Conclusions Cross theoretical model has good effect on the application ofFiveemergency technology training in the high altitude troops battlefield, and can effectively improve the troops self- help and mutual aid capabilities, it is worthy of popularization and application.

Objective To explore the effect of trans theoretical model in changing on healthy behavior of soldiers on plateau area to prevent hemorrhoids. Methods Fifty-four soldiers respectively from the artillery troops and air defenses force troops were selected and divided into control group that received normal healthy education and treatment group that received the healthy behavior intervene based on transtheoretical model and the changes of the behavior at each stage of intervention were observed and contrasted between two groups. Results There was no statistically significant difference in the number of each stage between two groups at 1 and 3 months after intervention (χ2= 1.514, 6.554, P> 0.05). The number of former intention stage, intention stage, preparation stage, action stage, maintain stage at 6 months after intervention in treatment group were respectively 0, 0, 0, 15, 39 cases, and 2, 3, 6, 20, 23 cases in control group respectively, the difference was statistically significant (χ2=15.843, P<0.01). There was no significant difference in the number of daily one form training at 1 months after intervention between the two groups (χ2=0.872, P>0.05). The number of daily training of the levator ani less than 5, 6-15, 16-25, 26-35,>35 times at 3, 6 months after intervention respectively was 5, 6, 6, 17 cases and 20, 0, 0, 0, 14, 40 cases in treatment group respectively, 9, 13, 14, 10, 8 cases, and 3, 4, 6, 22, 19 cases in control group,the differences were statistically significant (χ2=13.459, 22.252, P<0.01). The incidence of bloody at 6 months after intervention was 3.70% (2/54) in treatment group and 16.67% (9/54) in control group, the difference was statistically significant (χ2 = 4.960, P < 0.05). Conclusions The intervene based on transtheoretical model can effectively promote the healthy behavior to prevent hemorrhoids and reduce the recurrence rate of hemorrhoids.

Objective To establish a stable in vitro model of blood-brain barrier (BBB) simulating in vivo state using the primary-cultured rat brain microvascular endothelial cells (BMVECs) and pericytes.Methods The primary rat BMVECs and pericytes were isolated,purified and cultured.The isolated cells were identified by immunocytochemical staining method.An in vitro model of BBB was constructed using Transwell inserts (pore size 0.4 μm) coculture.Its barrier function was evaluated by the 4-hour leakage test,tight junction protein identification,transendothelial resistance detection,and permeability test.The difference between the cocultured model and simple BMVEC model across the membrane resistance values,and the permeability difference of the small molecule sodium fluorescein (Na-F) were compared.Results Confluent BMVEC monolayers demonstrated a typical cobblestone appearance and the pericytes displayed irregular shape and overlapping growth.Immunodouble labeling technique identification showed that the pericytes positively expressed α-srmooth muscle actin (α-SMA) and neuron-glial antigen 2 (NG2); after the fusion of cocultured model endothelial cells,the surface leakage test became positive; immnocytochemical staining shows that a continuous and dense tight junction formed between the endothelial cells; compared to the BMVEC model,the transendothelial electrical resistance of the cocultured model increased significantly (190.762 ± 10.326 Ω/cm2 vs.96.503 ± 8.012 Ω/cm2; t=- 24.489,P ＜0.01),and the permeability decreased significantly (56.149％ ± 3.572％ of the single endothelial model; t =19.330,P ＜ 0.01 ).Conclusions The primary isolated rat BMVECs and pericytes cocultured the morphology,structure and barrier function of in vitro model have the basic characteristics of BBB,and they have provided a useful tool for the research of BBB.

ObjectiveTo investigate the protective effect of paraoxonase 1 (PON1) gene against liver oxidative stress injury in mice due to dichlorvos poisoning.Methods Experiment 1: 12 male Balb/c mice were randomly divided into three groups, with 4 mice in each group: control group, green fluorescent protein lentivirus control group (Lv-GFP group), and recombinant PON1 lentivirus group (Lv-PON1 group). 2×107 TU of Lv-GFP or Lv-PON1 was transfected via tail vein, while normal saline was given to those in control group. Blood was collected on 0, 1, 3, 5, 7, 9 days via fundus venous plexus for the assay of serum PON1 activity. PON1 mRNA and protein expression levels were respectively determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot on the 3rd post-lentivirus transfection day. Experiment 2: according to the random number table method, another 96 male Balb/c mice were divided into four groups of 24 mice in each control group, dichlorvos group, Lv-GFP intervention group, and Lv-PON1 intervention group. Lv-GFP or Lv-PON1 was transfected via tail vein followed by intraperitoneal injection of dichlorvos 9 mg/kg, while those in control group were given normal saline. Six mice in each group were sacrificed respectively at 6, 12, 24, 48 hours, and liver tissue was collected. PON1 mRNA and nuclear factor E2-related factor 2 (Nrf2) mRNA expression levels were determined by RT-PCR, and PON1 protein level was determined by Western Blot. The content of malondialdehyde (MDA) and glutathione (GSH) in the liver tissue were determined by chemical colorimetry. The activity of superoxide dismutase (SOD) and catalase (CAT) were measured by double antibody sandwich enzyme linked immunosorbent assay (ELISA).Results Experiment 1: after Lv-PON1 was transfected to normal mice, PON1 activity in serum gradually increased and maintained a high level on 3rd day, while that of the control group and Lv-GFP group showed a normal low level. On the 3rd post-lentivirus transfection day, PON1 mRNA and PON1 protein expressions in liver were significantly higher than those of control group and Lv-GFP group. Experiment 2: compared with control group, the mice in dichlorvos group showed significant decreases in PON1 mRNA, PON1 protein, Nrf2 mRNA as well as GSH, SOD, CAT levels at 6 hours [PON1 mRNA (gray value):0.237±0.075 vs. 0.674±0.011, PON1 protein (gray value): 0.602±0.086 vs. 0.998±0.124, Nrf2 mRNA (gray value): 0.089±0.012 vs. 0.126±0.010, GSH (mg/g): 3.84±0.33 vs. 5.52±0.40, SOD (μg/g): 0.383±0.040 vs. 0.564±0.052, CAT (ng/g): 7.32±1.28 vs. 12.46±1.54, allP< 0.05], and remarkable increase in MDA content (nmol/g: 7.78±0.41 vs. 2.34±0.25,P< 0.05). With the extension of time, PON1 mRNA, PON1 protein, Nrf2 mRNA and GSH, SOD, CAT levels gradually increased, MDA content gradually decreased, Nrf2 mRNA expression level had risen to the level of control group at 24 hours (0.133±0.019 vs. 0.126±0.009,P> 0.05), and it was higher than that of the control group at 48 hours (0.206±0.028 vs. 0.124±0.010,P< 0.05). Compared with that of the dichlorvos group, Lv-PON1 intervention group showed a significant increase in PON1 mRNA, PON1 protein, Nrf2 mRNA and GSH, SOD, CAT levels [PON1 mRNA (gray value): 0.726±0.021 vs. 0.237±0.075, PON1 protein (gray value): 0.739±0.050 vs. 0.602±0.086, Nrf2 mRNA (gray value): 0.158±0.007 vs. 0.089±0.012, GSH (mg/g): 4.30±0.22 vs. 3.84±0.33, SOD (μg/g): 0.454±0.062 vs. 0.383±0.040, CAT (ng/g): 8.98±1.02 vs. 7.32±1.28, allP< 0.05], and a decrease in MDA content (nmol/g: 6.56±0.44 vs. 7.78±0.41,P< 0.05).Conclusion Regulation of PON1 gene can reduce MDA content, enhance SOD and CAT activities, increase GSH content, and it may also up-regulate Nrf2 mRNA expression to play a protective effect against oxidative stress of liver injury induced by dichlorvos poisoning.