Objective To study the effects of short-term and long-term exposure to extremely low frequency electromagnetic field(ELFEMF)on Morris learning and memory among rats. Methods Both younger(2-3 months old)and adult(18 months old)male SD rats were exposed to 50 Hz uniform ELFEMF with continuously adjusted intensities(0.1,1.2,1.6 mT) developed by Helmholtz coil pair for 10 days to 8 months. The levels of escape latency and the crossing annulus coefficients reflecting the behavioral changes in place navigation ability and spatial probe ability were examined by Morris water maze after exposure to ELFEMF among rats. Results The escape latency was shortened and the crossing annulus coefficients were increased compared with those of control group when the younger rats were exposed to 0.1,1.2 and 1.6 mT ELFEMF for 10 days respectively,and only revealed significant differences in 1.2 and 1.6 mT ELFEMF exposure groups compared with those of control group. In the younger rats of 1.6 mT ELFEMF exposure group with 10-day exposure ,their escape latencies were prolonged and the crossing annulus coefficients were decreased gradually and showed no significant differences compared with those of control group after the 30-day blank exposure period without ELFEMF.The younger rats of 1.2 mT ELFEMF exposure group showed significantly longer escape latency and lower crossing annulus coefficient compared those of control group after continuous 90-day exposure to 1.2 mT ELFEMF. The escape latency of adult rats in 0.1 mT ELFEMF exposure group showed no significant difference after 10-day exposure,was prolonged after 4-month exposure,and was significantly prolonged further more after 8-month exposure. Conclusion The learning and memory ability could be promoted by short-term ELFEMF exposure and impaired by long-term ELFEMF exposure for the young rats,and could be obviously inhibited by long-term ELFEMF exposure for the adult rats.

Objective To study the impact and mechanism of continuous venovenous hemofiltration (CVVH) in different uhrafihration rates on plasma cytokines in porcine endotoxemic shock. Methods Eighteen anesthetized mechanically ventilated pigs weighing 21-34 kg were randomly divided into three groups. In control group (n=6), the pigs received a 15.7 μg/kg endotoxin (E.coli 0111:84) infusion. In CVVH group (n=6) and high volume hemofihration (HVHF) group (n=6), the pigs received CVVH after the endotoxin infusion for 24 hours with an was taken before endotoxin infusion and at 0, 1, 6, 12, 24 h during CVVH. The plasma levels of TNF-α, IL-6, IL-10 and IL-18 were tested by ELISA. Results The survival time in control group was (15.4±5.2) h,CVVH group was (21.4±7.1) h,HVHF group was (22.4±6.7) h. The survival time in CVVH and HVHF group was significantly longer than that of control group (P< 0.05 ). Heart rate (HR), mean arterial blood pressure (MAP), central venous pressure (CVP) and cardiac output (CO) showed no significant differences among three groups. Plasma BUN and Ser increased gradually after the establishment of porcine endotoxemic shock model. BUN and Scr of CVVH and HVHF group were lower compared to control group (P<0.05), but there was no significant difference between CVVH and HVHF group (P>0.05). Plasma TNF-α and IL-6 peaked at T1, IL-10 peaked at TO, then they declined gradually. While IL-18 increased at TO and did not change after TO. A significant decrease of plasma IL-10 level was observed at T6, T12 and T24 in CVVH group compared with control group (P<0.05). HVHF group accomplished a greater decrease in plasma TNF-α (T6) and IL-10 (T6, T12, T24) levels compared with control group and CVVH group (P< 0.05). The levels of IL-6 and IL-18 showed no significant differences among three groups. There was a negative correlation between IL-6 and survival time (P<0.05). Conclusions HVHF and CVVH can prolong the survival time of porcine endotoxemic shock. IL-10 can be removed effectively with CVVH and HVHF. HVHF can also remove TNF-α effectively. CVVH and HVHF treatment can both remove BUN and Scr effectively. IL-6 is a powerful independent predictive factor for survival time of porcine endotoxemic shock.

Objective To investigate the preventive effect of somatostatin and nitroglycerin on post-ERCP pancreatitis (PEP) and hyperamylasemia.Methods A total of 184 patients who underwent endoscopic retrograde cholangiopancreatograph (ERCP) were enrolled,and randomly divided into three groups to receive somatostatin or nitroglycerin or normal saline according to random number table.Incidence of PEP and hyperamylasemia were observed and compared among the three groups.Results Nine patients stopped the medication and dropped out of the study after occurence of adverse reactions from receiving nitroglycerin injections.The incidence rates of hyperamylasemia were 38.7％ (24/62),51.8％ (28/54) and 37.3％ (22/59) in somatostatin,nitroglycerin and control group respectively,with no significant differences among the three groups (P ＞ 0.05).The incidence rates of PEP were 6.4％ (4/62),9.3 ％ (5/54) and 23.7％ (14/59) in somatostatin,nitroglycerin and control group,respectively.The incidence rates of PEP in somatostatin and nitroglycerin group were both significantly lower than that in control group (somatostatin group vs control group:x2 =7.13,P =0.01 ; nitroglycerin group vs control group:x2 =4.22,P =0.03).There was no significant difference between somatostatin group and nitroglycerin group (x2 =0.32,P =0.41).Conclusion Somatostatin and nitroglycerin injections after ERCP show similar efficacy on preventing PEP,but the former is safer.Both have little effects on the prophylaxis of hyperamylaemia.

Objective To investigate the safety,efficacy of CO2 insufflations during ERCP.Methods One hundred and forty eligible patients who underwent ERCP in Fuzhou General Hospital of Nanjing Military Command from January to December 2012 were randomized according to random digits in a double blind manner to receive either CO2 insufflations or air insufflations (65 vs 75).The patient's experience of abdominal pain and distension at 1 h,2 h,6 h and 24 h post-ERCP were evaluated by visual analogue scale (VAS).The heart rates,blood pressure,SpO2,PaCO2 were recorded before,during,after ERCP,and 24 h after the examination.Plain abdominal radiographic images were collected to show abdominal distention before and after ERCP,waist circumference,operative time,recovery time,intraoperative and post-ERCP complications were also determined.Results Post-ERCP the VAS of abdominal pain at 3 h was (4.08 ± 1.36) in CO2 group,which was significantly lower than that in air group [(4.59 ± 1.66),P ＜ 0.01].The VAS of abdominal distension at 1 h,3 h,6 h after ERCP were(1.78 ± 1.90),(0.72 ± 1.15),(0.12 ±0.45),which were significantly lower than those in air group [(3.53 ± 2.71),(2.51 ± 2.04),(0.84 ± 1.24),P ＜ 0.05] ; the intraoperative SpO2 was (93.29 ± 1.40)％,which was significantly lower than that in air group [(93.52 ± 1.06) ％,P ＜ 0.01].The waist circumference at 1 h after ERCP increased by (2.48 ± 1.33)cm,which was significantly lower than that in air group [(3.56 ± 2.13) cm,P =0.00).Recovery time was (11.2 ± 2.5) rain in CO2 group,which was significantly longer than that in air group [(9.7 ± 1.7) min,P =0.00].And the difference between the two groups was statistically significant.The plain abdominal radiographic images showed the degree of bowel distension was severer in air group than that in CO2 group (P =0.04).Conclusions CO2 insufflations are safe and efficacy during ERCP.CO2 can better alleviate abdominal pain and distension than air.

Objective To analyze proliferation and differentiation of glial fibrillary acid protein (GFAP)- and nestin-positive (GFAP+/nestin+) cells isolated from the subventricular zone following fluid percussion brain injury to determine whether GFAP+/nestin+ cells exhibit characteristics of neural stem cells. Methods Male Sprague-Dawley rats, aged 12 weeks and weighing 200-250 g, were randomly and evenly assigned to normal control group and model group. In the model group, a rat model of fluid percussion brain injury was established. Five days later, subventricular zone tissue was resected from each group and made into single cell suspension. After serum-free neural stem cell medium culture and subsequent serum-induced differentiation, cell type, proliferation and differentiation capacities were determined by immunofluorescence staining and flow cytometry. Results At 3-7 days after fluid percussion brain injury, nestin+/GFAP+ cells in the single cell suspension from the model group significantly outnumbered those from the normal control group (P<0.01). In the model group, an increased number of small neurospheres with smooth cell edge and bulged center formed after primary culture, and were clearly visible with the increase of culture time and medium replacement. After several passages, many clonal spheres were obtained, suggesting strong self-proliferatiing capacity. Neurospheres from the model group differentiated into astrocytes, neurons and oligodendrocytes. Conclusion GFAP+/nestin+ cells isolated from the adult rat subventricular zone after fluid percussion brain injury are thought to be neural stem cells because of their self-renewal and multi-differentiation capacities.

Thirty seven patients with congestive heart failure (CHF) were divided into cilazapril group (n=19) and general treatment group (n=18). Serum levels of interleukin-6 (IL-6) ,interleukin-8 (IL-8) , interleukin-10(IL-10) and tumor necrosis factor-α(TNF-α) , left ventricular ejection fraction (LVEF) ,left ventricular end-diastolic diameter (LVEDD), cardiac output (CO) and fractional shortening (FS) were measured before and after treatment. Serum levels of cytokines were also measured in 40 healthy individuals (control group). Results: The serum levels of IL-6, IL-8, IL-10 and TNF-α in CHF patients were significantly higher than those in the control group ( all P<0.01 ) ; After treatment, the serum IL-6, IL-8 and TNF-α were significantly decreased (P<0.01 ,P<0.05 ) in the cilazapril group. The LVEF, FS, CO were significantly increased in the Cilazapril group ( P<0.01 ) ; And the serum levels of IL-6 were significantly decreased in the cilazapril group as compared with the general treatment group ( P<0.05 ), however, after treatment, the EF, LVEF, FS and CO had no statistical differences in the cilazapril group as compared with the general treatment group. In the control group only LVEF and FS improved(P<0.01) ; Cytokine levels showed no changes. It suggests that cilazapril can reduce the serum levels of pro-inflammatory cytokines, increased the serum levels of anti-inflammatory cytokine, protect and improved cardiac function in the patients with congestive heart failure.

Objective To investigate the effects of repeated low dose intravenous infusion of low molecular weight iron dextran and iron sucrose on oxidative stress in chronic renal failure (CRF) rats. Methods CRF model was established by 5/6 subtotal nephrectomy (5/6 Nx). Four weeks after removing the right kidney, successful rats were randomly divided into low molecular weight iron dextran group, sucrose iron group and CRF control group. The sham group was established simultaneously. The dose of iron administrated in each rat was similar in iron dextran group and sucrose iron group. There were 6 rats in each group. Animals were observed for 6weeks, then the blood, urine and renal tissue samples were collected, and indexes of renal function,anemia, iron status and oxidative stress were investigated. Results The hemoglobulin (Hb) level in iron groups was significantly higher as compared to control group (P＜0.05) but was not significantly different between two iron groups. The levels of serum iron, ferritin and saturation rate of transferring (TS) were obviously lower in control group as compared to sham group (P＜0.05).Levels of above 3 indexes were significantly higher in two iron groups as compared to control group (P＜0.05), but were not significantly different between two iron groups. Concentration of plasma advanced oxidation protein products (AOPP) was obviously higher in two iron groups than that in control group [(127.84±21.19) μmol/L, (134.21±29.38) μmol/L vs (81.83±19.93) μmol/L, P＜0.05]. Plasma malonaldehyde (MDA) was significantly higher in iron sucrose group than that in iron dextran group [(6.06±0.73) nmol/L vs (4.99i0.80) nmol/L, P＜0.05]. Serum levels of superoxide dismutase (SOD) and total anti-oxidant capacity (TAOC) had no significant differences among three CRF groups. Concentration of plasma glutathione peroxidase (GSH-Px) was significantly decreased in three CRF groups as compared to sham group (P＜0.05), while plasma GSH-Px was significantly lower in sucrose iron group than that in iron dextran group and control group [(2123.11±74.78)nmol ·ml-1 ·min-1 vs (2352.84±163.90) nmol· ml-1 ·min-1, (2310.23±125.99) nmol ·ml-1 ·min-1, P＜0.05]. Conclusions Injection of intravenous iron can partially improve the anemia and the iron status indexes in 5/6 Nx CRF rats. Repeated low dose intravenous infusion of iron dextran and iron sucrose can aggravate the oxidative stress state in CRF rats, and the iron sucrose is worst.

In the present study, packaging system composed of pAAV-CMV-GFP, pAAV-RC and pHelper were transfected into human embryonic kidney 293 cells (HEK293 cells) mediated by polyethyleneimine (PEI) to explore an optimal transfection condition. Different total plasmid DNA dosages (1, 2, 3, 4, 5, 6 μg) and different PEI/Plasmid ratios (1:1, 3:1, 5:1, 7:1) were tested with detection of green fluorescence protein (GFP) with ImagePro Plus6. 0 Software. Then transfection efficiency of the optimized transfection system was further observed for different time periods(12, 24, 36, 48, 60, 72 h). The results showed that total plasmid dosage of 4 μg/well with PEI/plasmid ratio of 3 : 1-5 : 1 was an efficient transfection condition. Transfection efficiency-time curve was an S-shaped curve. Transfection efficiency reached a plateau at 60 h after transfection. The optimized conditions for PEI-mediated transfection at the optimal time result in enhanced transfection efficiency of triple plasmid into HEK293 cells.
Green Fluorescent Proteins ; HEK293 Cells ; Humans ; Plasmids ; Polyethyleneimine ; Transfection ; methods

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ObjectiveTo study the anti-inflammatory effect of traditional Chinese medicine (TCM) Changweishu on sepsis patients with gastrointestinal dysfunction and its protective effect on intestinal mechanical barrier.Methods A prospective research method was conducted. Fifty sepsis patients with gastrointestinal dysfunction admitted into Department of Emergency Intensive Care Unit (EICU) of Hebei Provincial TCM Hospital from October 2013 to June 2015 were enrolled, and they were divided into a conventional treatment of western medicine control group and a TCM Changweishu group according to the random number table method, 25 cases in each group. The conventional western medicine treatment was given to both groups, and in TCM Changweishu group, additionally Changweishu 1 dose per day, a mixture of following ingredients constituting one dose without decoction, was applied, including: rhubarb 9 g, dandelion 20 g, green tangerine peel 15 g, angelica sinensis radix 15 g, red and white peony each 12 g, rhizome ligusticum 9 g, agrimony 20 g, coptidis rhizoma 6 g, rhizoma pinelliae 6 g, fructus trichosanthis 15 g, garden burnet root 20 g, poria 20 g, and taken once 1/2 dose in the morning and once 1/2 dose in the evening by oral or nasal feeding. After treatment for 7 days, the level changes of inflammatory factors of tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6), high mobility group protein B1 (HMGB1) and gastrointestinal dysfunction indexes of diamine oxidase (DAO), D-lactic acid in the two groups were observed.Results After treatment, the levels of TNF-α, IL-6, HMGB-1, DAO, D-lactic acid in the two groups were significantly lower than those before treatment [the western medicine control group: TNF-α (A value) 10.89±1.65 vs. 50.30± 1.58, IL-6 (A value) 7.35±1.85 vs. 12.66 ± 1.43, HMGB1 (A value) 5.28 ±1.64 vs. 6.23±0.95, DAO (A value) 5.87±0.59 vs. 6.67±0.49, D-lactic acid (A value) was 2.63±0.17 vs. 4.58±0.16; TCM changweishu group: TNF -α (A value) 5.38±1.19 vs. 51.23±2.34, IL-6 (A value) 5.54±1.26 vs. 13.24±1.78, HMGB1 (A value) 3.69±1.09 vs. 6.14±1.42, DAO (A value) was 3.39± 0.40 vs. 6.70±0.34, D-lactic acid (A value) 1.95±0.13 vs. 4.63±0.11, allP < 0.05]. After treatment, the degrees of decline in indexes of TCM Changweishu group were more obvious than those in western medicine control group [TNF-α(A value) 5.38±1.19 vs. 10.89±1.65, IL-6 (A value) 5.54±1.26 vs. 7.35±1.85, HMGB1 (A value) 3.69 ±1.09 vs.5.28±1.64, DAO (A value) 3.93±0.40 vs. 5.87±0.59, D-lactic acid (A value) 1.95±0.13 vs. 2.63±0.17, allP <0.05].Conclusions TCM Changweishu has protective effect on sepsis patients with gastrointestinal dysfunction, and its mechanisms are the amelioration of damage in intestinal tract mechanical barrier, decrease of permeability of intestinal mucosa and inhibition of levels of inflammatory factors.