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 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 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 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.

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.

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

Objective To observe the effect of heated humidified high flow nasal cannula oxygen therapy (HFNC) on patients with post-stroke systemic inflammatory response syndrome (SIRS). Methods Totally 78 patients with post-stroke SIRS were selected in the department of neurology of Wuxi People's Hospital and were randomly divided into HFNC group (n=40) and conventional therapy group (n = 38). The neurological impairment score (NIHSS) , APACHE-Ⅱ, clinical pulmonary infection score (CPIS) , C-reactive protein (CRP) of the 2 groups were recorded before and after the treatment. At the same time, modified Rankin score (mRS) of the two groups were also recorded. Results There was no significant difference in terms of morality and the number of patients with mechanical ventilation in the 2 groups. The 7-day APACHE Ⅱ, 7-day CPIS, 7-day SIRS cure rate, 14-day NIHSS and mRS of 3 months in HFNC group were higher than those in the conventional therapy group (P < 0.05).There was no significant difference in 7-day CRP, 14-day CRP and 14-day CPIS between HFNC group and conventional oxygen therapy group (P> 0.05). Conclusions HFNC can improve lung infection of patients with SIRS thus improve the recovery rate of SIRS. At the same time, it can improve the recovery of the neurological deficit and prognosis in acute stroke.

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.

[ ABSTRACT] AIM:To investigate the role of ATP-sensitive potassium ( KATP ) channels in the inhibitory effect of hydrogen sulfide ( H2 S) on high glucose ( HG)-induced inflammation mediated by necroptosis in H 9c2 cardiac cells. METHODS:The expression levels of receptor-interacting protein 3 ( RIP3; an indicator of necroptosis ) and cyclooxyge-nase-2 (COX-2) were determined by Western blot.The levels of interleukin-1β(IL-1β) and tumor necrosis factor-α( TNF-α) were detected by ELISA .RESULTS:After H9c2 cardiac cells were treated with 35 mmol/L glucose ( HG) for 24 h, the expression of RIP3 was significantly increased .Pre-treatment of the cells with 100 μmol/L diazoxide ( DZ; a KATP channel opener) or 400 μmol/L NaHS (a donor of H2S) for 30 min considerably blocked the up-regulation of RIP3 induced by HG.Moreover, pre-treatment of the cells with 100 μmol/L 5-hydroxydecanoic acid (5-HD; a KATP channel
blocker) attenuated the inhibitory effect of NaHS on HG-induced up-regulation of RIP3.On the other hand, co-treatment of the cells with 100μmol/L necrostatin-1 ( a specific inhibitor of necroptosis ) or pre-treatment of the cells with 100 μmol/L DZ or 400 μmol/L NaHS attenuated HG-induced inflammatory responses , evidenced by decreases in the expression of COX-2 and secretion levels of IL-1βand TNF-α.However , pre-treatment of the cells with 100 μmol/L 5-HD significantly attenuated the above anti-inflammatory effects of NaHS.CONCLUSION:KATP channels play an important role in the inhib-itory effect of H2 S on HG-induced inflammation mediated by necroptosis in H 9c2 cardiac cells.

Objective To investigate the immunoregulatory effects of mesenchymal stem cells (MSCs) on the peripheral T-lymphocytes of lupus nephritis in vitro. Methods MSCs were isolated and expanded from human bone marrow cells. The purity of MSCs was identified by flow cytometry (FCM). The MSCs (4×104, 1×104, 2×103) were added into wells containing peripheral blood lymphocytes (2×105) from lupus nephritis in the presence of phytohemagglutinin [PHA). Samples were divided into the following groups: group A:T-lymphocytes alone; group B: MSCsl with T-lymphocytes(MSCsl:T=1:5); group C: MSCs2 with T-lymphocytes (MSCs2:T=1:20); group D: MSCs3 with T-lymphocytes (MSCs3:T=1:100). The proliferation of T-lymphocytes was assessed by MTT. FCM was used to analyze the apoptosis of T-lymphocytes and surface markers of CD28 and CD152. The gene expression of interferon γ (IFN-γ), interleukin-10 (IL-10), transforming growth factor-β1 (TGF-β1) were detected by real-time RT-PCR. Results In vitro, with the presence of MSCs, peripheral blood T-lymphocytes from lupus nephritis were statistically significantly decr-eased in their proliferative activities , apoptosis and CD28 expression in a dose-dependent manner. No inhibitory effects on CD152 expression of T-lymphocytes had been observed . MSCs promoted the gene expression of TGF-β1 and inhibited the gene expression of IL-10, IFN-γ. Conclusion MSCs can inhibit the proliferative activities, apoptosis and CD28 expression of peripheral blood T-lymphocytes of lupus nephritis, increase gene expression of TGF-β1 and lower the gene expression of IL-10, IFN-γ, which may play an important role in it's immunosuppressive effects on lupus nephritis.