RESUMO
Objective To explore the protective effects of adipose - derived stem cells (ADSCs) with phosphodiesterase 5 inhibition by lentivirus-mediated stable gene silencing on the proliferation and apoptosis of renal tubular epithelial cells induced by ischemia-reperfusion injury in vitro. Methods To isolate cultivate and indentify ADSCs from rats. Lentiviral expression vector of carrying PDE5 shRNA gene was transfected into ADSCs, and a negative control group was set up.Western blotting was used to detect PDE5 protein expression levels. ADSCs were co-cultured with NRK-52E in a transwell system, and NRK-52E cells were treated with ischemia/reoxygenation protocol. Edu assay was performed to evaluate the proliferation of NRK cells, flow cytometry to detect the apoptosis of NRK cells, and ELISA to quantify the protein expressions of fibroblast growth factor (FGF) and hepatocyte growth factor (HGF). The expression of E - cadherin and cytokeratin 18 (CK18) was quantified by real time PCR and flow cytometry. Results Western blotting for PDE5 protein indicated a significant reduction of PDE5 protein levels in PDE5 shRNA transduced population. After the treatment of ischemia/reoxygenation in vitro, the proliferative viability and apoptosis of NRK-52E cells co-cultured with ADSCs induced by PDE5 gene inhibition were significantly improved, compared to the normal group (all P<0.05). And the release of HGF, FGF were markedly enhanced (all P<0.05). Moreover, the NRK-52E cells survival, the expression of E-cadherin and CK18 on PDE5 inhibited ADSCs co-cultured with I/R injured NRK cells was significantly increased compared to that in the negative control group (all P<0.05). Conclusion ADSCs preconditioned by inhibition of PDE5 can be a powerful novel approach to improve the survival of renal tubular cells following ischemia-reperfusion injury, and have an obvious tendency to transform epithelial cells.
RESUMO
Objective To explore the in vitro effect of lentiviral expressions vector carrying phosphodiesterase 5 (PDE5)-shRNA gene on proliferation,apoptosis and secretion of adipose-derived stem cells (ADSCs).Methods (1) ADSCs were separated and purified from the rats,sub-cultivation and identification were performed.Lentiviral expression vectors carried different PDE5 shRNA genes (PDE5 shRNA1,PDE5 shRNA2 and PDE5 shRNA3) were established and negative controls (NC shRNA) were used.The above vectors were transfected into ADSCs with lentivims package;Western blotting was carried out to detect the PDE5 protein expression level and real time-PCR was carried out to detect the mRNA expression level so as to select the most efficient cell line.(2) ADSCs were divided into four groups:normal control group,ischemia-re-oxygenation (I/R) injury group (normal cells+I/R injury),NC group (NC shRNA-ADSCs+I/R injury) and experimental group (PDE5 shRNA-ADSCs+I/R injury);CCK-8 assay and Edu method were per;ormed to evaluate the proliferation of cells,flow cytometry was employed to detect the apoptosis of ADSCs,and ELISA was used to quantify the protein expressions of vascular endothelial growth factor (VEGF),basic fibroblast growth factor (b-FGF) and cyclic guanosine monophosphate (cGMP).Results Lentiviral expression vectors carried different PDE5 shRNA genes were successfully established and PDE5 sh ADSCs with stable PDE5 expression were established.As compared with cells from normal control group,cells from I/R injury group had significantly decreased ADSCs proliferation rate and increased apoptosis rate at early and late stages,and significantly increased VEGF and FGF levels in the liquid supernatant and cGMP level in the cells (P<0.05);these differences were not significant between the I/R injury group and NC group (P>0.05);as compared with the NC group,the experimental group had significantly increased ADSCs proliferation rate and decreased apoptosis rate,and significantly increased VEGF and FGF levels in the liquid supematant and cGMP level in the cells (P<0.05).Conclusion The proliferation,apoptosis and secretion of ADSCs could be effectively improved and the stem cell efficiency was modified via up-regulating cGMP level after PDE5 gene expression being inhibited in ADSCs.
RESUMO
Objective To investigate and compare the change of extravascular lung water (EVLW) and levels of cytokines in septic patients without clinical acute lung injury (ALI) /acute respiratory distress syndrome (ARDS) with those in spetic patients with sepsis-induced ALI/ARDS in order to determine the role of EVLW involved in the pathogenesis of lung injury in the patients by quantifying the relationship between EVLW and biomarkers of lung injury in patients with sepsis.Methods A total of 40 septic patients complicated either with or without clinical ALI/ARDS after sepsis. In each patient,transpulmonary thermodilution (PiCCO) was used to measure cardiovascular hemodynamics and EVLWI for 7 days via an arterial cannula indwelled within 72 hours after diagnosis of severe sepsis was made, and serial bronchoalveolar lavages (BAL) were carried out.Other examinations including blood gas analysis,ventilator parameters,chest X-ray and the levels of pro-inflammatory cytokines such as tumor necrosis factor (TNF-oα),interleukin-1 in the BAL were recorded.In-hospital and ICU mortalities were also observed.Results Of total 40 patients,29 were complicated with clinically defined septic ALI/ARDS ( ARDS n =15,and ALI n =14).The septic patients complicated with ALL/ARDS had significantly higher amount of EVLWI and higher levels of TNF-α and interleukin-1 in the BAL than patients without ALI/ARDS ( P < 0.05).The arterial oxygen tension/fractional inspired oxygen ratio,lung injury score,and the levels of TNF-α,IL-1 in the BAL correlated with EVLWI.Moreover,in-hospital mortality,ICU mortality and the length of ICU stay of the patients with high amount of EVLWI were markedly increased than those of patients with low amount of EVLWI. Conclusions In septic patients complicated with ALI/ARDS, the extravascular lung water index correlates with oxygenation,lung injury severity and inflammatory cytokines in lung.Determination of EVLWI may be useful for evaluation of severity of lung injury and prognosis of septic patients.
RESUMO
ObjectiveTo evaluate the effect of xuebijing injection on inflammatory response and cellular immune function in patients with severe sepsis.MethodsSixty-two patients with severe sepsis from September 2008 to August 2009 were divided into treatment group(30 patients) and control group (32patients) by random digits table.All the patients received sepsis-bundle therapy and patients in treatment group added xuebijing injection therapy with 100 ml,twice a day,for 7 days.The levels of serum tumor necrosis factor-alpha(TNF-α ),interleukin(IL)-6,IL-10 and C-reactive protein (CRP),peripheral blood T lymphocyte CD4+、CD8+ 、CD4+/CD8+,the expression of human leucocyte antigen (HLA)-DR on CD14+peripheral blood mononuclear cell (PBMC) were detected before and aftertreatment.ResultsThere was no significant difference in the levels of serum TNF-o,IL-6,IL-10,CRP,peripheral blood Tlymphocyte CD4+、CD8+、CD4+/CD8+ and the expression of HLA-DR on CD14+ PBMC before treatment between two groups(P > 0.05).After treatment,compared with those in control group,the levels of serum TNF- α,IL-6,IL-10 and CRP in treatment group were significantly decreased [ ( 64.4 ± 13.5) ng/L vs.(96.1 ± 22.1 ) ng/L,( 153.8 ± 23.8 ) ng/L vs.(180.1 ± 21.7) ng/L,(73.8 ± 13.8) ng/L vs.(101.1 ± 11.7) ng/L,(53.7 ± 18.8) mg/L vs.(91.3 ± 32.8)mg/L,P <0.05],while peripheral blood T lymphocyte CD4+/CD8+ and the expression of HLA-DR on CD14+PBMC were significantly increased [ 0.311 ± 0.021 vs.0.424 ± 0.035,0.201 ± 0.017 vs.0.238 ± 0.038,1.78 ±0.21 vs.1.56 ±0.18,(38.4 ± 11.5)% vs.(18.1 ± 12.1)%,P<0.05].ConclusionXuebijing injection can reduce the inflammatory response and ameliorate immune disorder in patients with severe sepsis.
RESUMO
Objective To study the protective effect of removing inflammatory cytokines by hemoperfusion (HP)on acute kidney injury (AKI) in patients with sepsis. Method A total of 40 patients with sepsis and AKI were randomly divided into two croups: HP treatment group (n = 22) and control group (n = 18). Hemoperfusion carried out in patients of Hp group with HA330 filter once a day for 3 days and the procedure of each hemoperfusion was completed in 2 hours. The patients of control group were treated with routine treatment. Further, the hemodynamics, plasma IL-6, IL-10, C-reactive protein (CRP), serum creatinine (Scr), blood BUN and urine NAG, γ-GTP,α1-MG of patients in both groups were detected before treatment and 3 d,7 d and 14 days after treatment. Results Compared to control group, the levels of plasma interleukins-6, IL-10 and C-reactive protein were significantly lower (P < 0.05), along with increase in urine output, lower levels of blood BUN and Scr, reduction in urine NAG,γ-GT and α1-MG (P < 0.05). In addition, the patients at Ⅰ or Ⅱ stage of AKI treated with hemoperfusion had significantly lower level of Scr in 14 days and lower mortality in intensive care unit in comparison with control group (P < 0.05). Conclusions Hemoperfusion employed in the earlier stage of AKI with the HA type filter may have protective effect on acute kidney injury by the removal of inflammatory cytokines in the setting of sepsis.
RESUMO
Objective To analyze the alterative characteristics of the extravascular lung water (EVLW ) in the patients with septic shock and clarify its value on the prognosis of these patients.Methods By the methods of retrospective analysis,according to the ultimate survival,21 patients with septic shock were divided into survivor group (10 cases) and non-survivor group (11 cases).The clinical features of the patients were observed and hemodynamic monitoring was made with PiCCO monitor.The EVLW was measured and the relationship between the EVLW and the prognosis of patients was analyzed.Results On the first,second and third day,EVLW was (12.7 ±1.8),(11.3 ±1.3),(10.1 ±1.3) ml/kg in survivor group,and (14.4 ± 1.0),(14.6 ± 1.4),(14.6 ±1.3) ml/kg in non-survivor group respectively,and there were statistical differences between two groups (P 0.05).Conclusions The EVLW in the patients with septic shock increases significantly.The dynamic changes of the EVLW may be one of the factors for predicting the prognosis of patients with septic shock.
RESUMO
Objective To explore the effect of mild hypothermia on inflammation status,lung function protection and clinical prognosis in patients with acute respiratory distress syndrome (ARDS).Methods All of 56 patients with ARDS were randomly divided into two groups: trial group (29 patients,treatment with mild hypothermia) and control group (27 patients, treatment with common practice). The following parameters including tumor necrosis factor (TNF)-α,interleukin (IL)-6 and C reactive protein (CRP), oxygenation index, SOFA evaluation and injury of lungs evaluation were detemined before treatment and at the 3rd, 7th day after treatment, and survival rates and adverse reaction in 28 days also were observed.Results After treatment, the levels of TNF-α ,IL-6 and CRP were decreased significantly, and oxygenation index, the scores of SOFA evaluation and injury of lungs evaluation were improved significantly in trial group than those in control group (P<0.05 ). The survival rate in trial group was higher than that in control group after treatment of 28 days [65.5%(19/29) vs 51.9%(14/27)]. The courses of mechanical ventilation and staying in ICU in trial group were shorter than those in control group [(11.9±3.6)d vs (17.0±5.1)d,(14.1±4.2)d vs (21.5±7.7)d](P<0.05). Conclusion Mild hypothermia can effectively attenuate inflammation disorder, improve damaged lung function and prognosis in patients with ARDS.
RESUMO
Objective To explore the protective effect of proliferator activated receptor-γ (PPAR-γ)activator pioglitazone on the expression of inflammatory cytokines in cultured cortical neurons after ischemia-reperfusion injury and its mechanism. Method The ischemie-reperfusion model was established by deprivating both glucose an oxygen in medium and then gave them back. Medium or that with pioglitazone was added at the beginning of reperfusion. The MTT values of neurons were determined in control or treatment groups, ANOVA was used to detect the expression of PPAR-γ. The expression of tumor necrosis factors-α(TNF-α) and interleukin-lβ(IL-lβ) were detected by Western Blotting. Results Compared to control group, the markedly reduction of MTT values and enhanced expression of PPAR-γ, TNF-a and IL-1β was observed in the ischemia-reperfusion neurons (P < 0.05). After they were treated by pioglitazone, the reduction of MTT values and enhanced expression of TNF-a and IL-1β were prominently reversed by the further activation of PPAR-γ ( P < 0.05). Conclusions Treatment of PPAR-γ activator pioglitazone has protective effect on neurons after ischemia-reperfusion injury. Its mechanism may be associated with the inhibition of inflammation after injury.
RESUMO
Objective To study the effect of continuous blood purification on inflammation and nutritional status in patients with sepsis, and to study relationship between inflammatory, malnutrition and illness. Method Forty-eight patients with severe sepsis were randomly divided into two groups: continuous renal replacement treat-ment (CRRT) group (n=27) and control group (n=21). The flowing biomarkers including albumin, preallbu-min, transfenin, insulin-like growth factor-1 (IGF-1), tumor necrosis factor (TNF-α) and C reactive protein (CRP) were determined before,and 1 week and 2 weeks after treatment. Results Compared with control group, levels of albumin, prealbumin and IGF-1 in treatment group increased significantly at 14 days after CRRT (P<0.05), and levels d CRP and TNF-α decreased significantly (P<0.05), resulted in increase in survival rate, shortened me-chanical ventilation time and decreased SOFA scores (P<0.05). The IGF-1 and prealbumin had a negative cor-relations with CRP and TNF-a in both groups (P<0.05). The survivors in both groups had significantly higher levels of IGF-1,prealbumin and TNF-α than the deads before trearment(P<0.05). Conclusions Continuous blood purification can effectively attenuate inflammation and improve nutritional status in patients with severe sep-sis.Maybe IGF-1 and prealbumin act as prognostic markers more sensitive in severe sepsis.
RESUMO
Objective To evaluate the clinical effect of ambulatory labor analgesia used in latent phase of the first stage of labor, which include labor progress, Apgar score after ambulatory labor analgesia begun to use when the cervix was different size dilatated. Methods Seventy-five parturient primiparas who had no complication were randomly divided into three groups: group Ⅰ: ambulatory labor analgesia was begun to use when the cervix was 1.0 cm dilated, group Ⅱ: ambulatory labor analgesia was begun to use when the cervix was 2-3 cm dilated, group Ⅲ: control group without use of ambulatory labor analgesia. Analgesic effects were observed, changes of uterine contraction were recorded by fetal monitor. Meanwhile, total stage of labor ,outcome of delivery and Apgar score were recorded. Results Duration of total stage of labor had no significant difference between group Ⅰ and group Ⅲ.The duration of the first labor stage was significantly longer in group Ⅰ than that in group Ⅱ(P
RESUMO
Objective: To evaluate the feasibility and safety of postoperative patient-controlled epidural analgesia (PCEA)in children. Methods: Forty postoperative pediatric patients(5-11 years old)were divided into two groups. A and B. Both wereinstituted with postoperative PCEA with LCP model (loading dose 2.05 + 0.13ml), continuos infusion rate 0.82 + 0.15mi/h, PCA dose 0.81 + 0.16ml)by Graseby-9300 PCA pump. The PCEA solution of group A was 0.075% bupivacaine plus0.0012 % buprenorphine, that of group B was same while 0. 005% droperidol was added as an adjuvant. Results:The volumeof PCEA sdution consumption in group B was significantly less than that in group A on the first and second postoperative day (P< 0.01-0.05). Good analgesic efficiency with little side effects was obtained, as evaluated by the VASF emasay or D/D score and complications in two groups, but the analgesic effect of group B was better than that of group A. Conclusion: Thepediatric PCEA with low concentration of bupivaeaine plus buprenorphine is feasible and safe. Droperidol may enhance theanalgesic effects of PCEA.
RESUMO
Objective To compare the safety and efficacy of different doses of intrathecal sufentanil for labor analgesia in parturients. Methods One-hundred ASA I or II nulliparous parturients at 39-40 weeks of gestation who requested labor analgesia were included in this randomized double-blinded study. They were all in active labor with a 2-3 cm cervical dilatation. The subarachnoid block was performed at L2, 3 . The patients were randomized to received intrathecal sufentanil 1,3,5,7,or 10 ?g. An epidural cather was then placed for patient-controlled epidural analgesia (PCEA) . The PCEA solution contained 0.1% ropivacaine and sufentanil 0.4 ?g ? kg-1 .The PCEA settings were as follows: background infusion 5 ml?h-1 , demand bolus 3 ml and lock-time interval 10min. Pain relief was assessed by VAS scores, oxytocin dose, maternal satisfaction, hemodynamics and side effects. Apgar scores of the new-borns were recorded. Results Demographic data including age, sex, body weight and baseline VAS were similar among the five dose groups. There were significant differences in the onset , duration and efficacy of analgesia among the five dose groups. Significantly more patients who received 10 ?g sufentanil reported nausea and vomiting and pruritus than those who received 1 or 3 fig sufentanil. There was no significant difference in Apgar scores among the 5 dose groups. Conclusion Intrathecal sufentanil combined with PECA is safe and effective for labor analgesia. Intrathecal sufentanil 3-5 ?g is the dose of choice for labor pain relief.
RESUMO
To determine the differences in analgesic, hemodynamic and ventilatory effects between epidural and intravenous midazolam (MID), 44 adult patients, ASA grade Ⅰ to Ⅱ, scheduled for elective upper abdominal surgery. were randomly allocated to receiving epidural MID 0.05 mg/kg (group E_1) or 0.1 mg/kg (group E_2). or intravenous MID 0.05 mg/kg (group V_1) or 0.1 mg/kg (group V_2), respectively, The analegic effect was evaluated by pin-prink test, following MID administration. The hemodynamie and ventilatory values were measured by impedance cardiogra phy and side stream spirometry, before and 1.3, 5, 10, 15 and 20 mins after MID administration. Tbe results showed that the segmental spinal analgesia occured with MID in group E_1 and E_2, but no analgesia was produced with WID in group V_1 and V_2. Following MID administration in four groups, HR increased during first 5 mins (P0.05); CI, SI. LVWI, MAP, RPP and indices of myocardial eonstraction, cardiae peak flow and blood ejection velocity decreased slightly(P0.05). After MID administration, SpO_2 and VT were reduced (P0.05), and the inspiratory end-tidal oxygen content difference was elevated transiently (P0.05) and went down in group E_2 and V2(P0.05).
RESUMO
Objective: To compare the clinical effects of postoperative patient controlled epidural analgesia (PCEA) with different combinations of tramadol and/or buprenorphine. Method:The 150 cases of surgical patients were ran domly divided into 6 groups: 0.6% tramadol (group T_1). mixture of 0.6% tramadol and 0.15% bupivacaine (group T_2), mixture of 0.0015% buprenorphine and bupivacaine (group B_1) , mixture of 0. 0015% buprenorphine, bupivacaine and 0.6% tramadol(group B_2). 0.15% bupivacaine(group C_1), 0.0015% buprenorphine(group C_2). With a Graseby- 9300 PCA pump in the loading-continuous-PCA model, the doses were 5mL-0.5ml-1ml respectively. Result: (1)In the first postoperative 24 hours the amount of drugs used in each group was similar; (2)1n these six groups, both visual analog scale scores and the ratios of tatol deliveries to demands were B_2
RESUMO
Objective To realize the hygienic state of drinking water in the rural areas in Yunnan Province,the scientific evidences would be provided for drawing the policies about improving the drinking water quality in the rural areas.Methods From August to October in 2006,30 counties were randomly chosen among the monitoring counties,in each county at least 10 sampling points were chosen randomly.The questionnaires were employed and the water samples were collected,the quality of drinking water was tested with the related standard methods.Results 8.94 million people were involved in the survey,48.60% of them used surface water as the source of water.52.18% of people were serviced by the central water supply.The eligible rate in 301 water samples in this survey was 36.88%,those of the samples of surface and underground water were 37.60% and 36.36% respectively.The eligible rate of samples from central water supply was 41.85%,higher than that from non-central water supply(P