AIM: To investigate whether Janus kinase/signal transducer and activator of transcription ( JAK/STAT) signaling pathway mediates high glucose-induced damage in human umbilical vein endothelial cells ( HUVECs ) . METHODS:The cell viability was examined by CCK-8 assay.The expression levels of JAK2, STAT3, caspase-9 and en-dothelial nitric oxide synthase ( eNOS) were detected by Western blot .The intracellular levels of reactive oxygen species ( ROS) were tested by DCFH-DA staining followed by photofluorography .Mitochondrial membrane potential ( MMP) was measured by rhodamine 123 staining followed by photofluorography .RESULTS:Treatment of HUVECs with high glucose (40 mmol/L glucose) for 6~12 h enhanced the protein level of phosphorylated JAK 2, peaking at 9 h.Treatment of the cells with high glucose for 6~12 h also increased the protein level of p-STAT3 with the peak value at 12 h.Pretreatment with the inhibitor of JAK/STAT pathway AG490 for 1 h before exposure of the HUVECs to high glucose significantly inhibi-ted the high glucose -induced injury , as evidenced by an increase in the cell viability , decreases in the expression of caspase-9 and the intracellular ROS production , and increases in MMP and the expression of eNOS .CONCLUSION:JAK/STAT signaling pathway is involved in the high glucose-induced damage in HUVECs .

AIM: To explore whether exogenous hydrogen sulfide (H2S) depresses high glucose (HG)-in-duced injury by modulating the Janus kinase/signal transducer and activator of transcription ( JAK/STAT) pathway in hu-man umbilical vein endothelial cells (HUVECs).METHODS:The protein levels of JAK2, STAT3 and cleaved caspase-3 were determined by Western blot.The cell viability was measured by CCK-8 assay.Mitochondrial membrane potential ( MMP) was detected by rhodamine 123 staining followed by photofluorography.The intracellular level of reactive oxygen species (ROS) was analyzed by DCFH-DA staining followed by photofluorography.The activity of superoxide dismutase (SOD) was also measured.RESULTS:Pretreatment of the HUVECs with 400 μmol/L NaHS (a donor of H2S) for 30 min prior to exposure to 40 mmol/L glucose ( HG) markedly attenuated HG-induced upregulation of the phosphorylation of JAK2 and STAT3.Pretreatment with 400μmol/L NaHS for 30 min or with 20μmol/L AG490 (inhibitor of the JAK/STAT pathway) for 30 min attenuated the injury of HUVECs induced by HG, as indicated by the increases in cell viability and SOD activity, and decreases in the protein level of cleaved caspase-3, ROS generation and dissipation of MMP.CONCLU-SION:Exogenous H2 S protects HUVECs against HG-induced injury by inhibiting JAK/STAT pathway.

AIM: To explore whether necroptosis contributes to the high glucose (HG)-induced damage in hu-man umbilical vein endothelial cells (HUVECs).METHODS: The protein levels of receptor-interacting protein 3 (RIP3) and cleaved caspase-3 were detected by Western blot.The intracellular levels of reactive oxygen species (ROS) were deter-mined by DCFH-DA staining followed by photofluorography.Mitochondrial membrane potential (MMP) was measured by rhodamine 123 staining followed by photofluorography.RESULTS: Treatment of HUVECs with HG at different concentra-tions (10, 20 and 40 mmol/L glucose) for 24 h gradually enhanced the expression levels of RIP3.Treatment of HUVECs with HG (40 mmol/L glucose) for different time (3 h, 6 h, 9 h, 12 h and 24 h) also up-regulated the expression levels of RIP3, peaking at 9 h.Pretreatment of HUVECs with 20 μmol/L Z-VAD-FMK (an inhibitor of caspase) for 30 min before exposure to HG enhanced the expression level of RIP3.Pretreatment of HUVECs with 100 μmol/L necrostatin-1 (an inhi-bitor of necroptosis) for 1 h before exposure to HG alleviated the HG-induced injuries, such as a decrease in cell viability, an increase in ROS generation and dissipation of MMP, but up-regulated the protein level of cleaved caspase-3.CON-
CLUSION: Necroptosis mediates HG-induced injury in HUVECs.There is a negative interacting between necroptosis and apoptosis.

ObjectiveTo observe the effects of norepinephrine and dobutamine (NE+Dobu) on hemodynamics, tissue perfusion and oxygen metabolism in elderly patients with septic shock at different levels of mean arterial pressure (MAP). MethodsAfter aggressive fluid resuscitation,norepinephrine and dobutamine were administered in 18 elderly patients with septic shock to correct hypotension. The rate of dobutamine infusion was 5 ug. kg<'-1>. min<'-1> Norepinephrine was titrated to maintain MAP at levels of 65 mm Hg, 75 mm Hg and 85 mmHg. Four hours later, the changes of hemodynamics, oxygenation index, blood lactate, renal function and gastric pCO<,2> at every MAP level were observed. ResultsThere were no significant differences in heart rate (HR), pulmonary arterial wedge pressure (PAWP), volume of oxygen (VO<,2>), Blood lactate, pH value, △pCO<'2> and creatinine clearance rate (CCr) among the MAP levels of 65 mm Hg, 75 mm Hg and 85 mm Hg ( all P>0.05). Compared with MAP 65 mm Hg, cardiac index (CI), systemic vascular resistance index (SVRI), oxygen delivery over oxygen extraction ratio were obviously higher at MAP 75 mm Hg and 85 mm Hg [4.7±0.6, 5.1±0.7 vs. 4.0±0.6; 1162±278, 1276±319 vs. 1011±225; (697±53) ml. min<'1>. m<'2>,(711±68)ml. min<'-1>. m<'2> vs. (634±70) ml · min<'-1> · m<'2>; (0.28±0.02)%,(0.27±0.02)% vs. (0.25±0.02) %, respectively, all P<0.05). The urinary output at MAP levels of 65 mm Hg, 75 mm Hg and 85 mm Hg were (98±43)ml/h, (91±54) ml/h and(74±49)ml/h repectively, and only the differences between MAP 75 mm Hg and MAP 65 mm Hg had statistical significance (P<0.05).ConclusionsAfter aggressive fluid resuscitation, hemodynamics and renal function are improved at MAP 75 mm Hg after administration of norepinephrine and dobutamine in elderly patients with septic shock. It should be considered that MAP is appropriately increased in elderly patients with septic shock.

Objective To explore the use of pulse indication contour cardiac output (PiCCO) as a guidance for fluid resuscitation and vasopressors employment in patients with septic shock in order to find out the efficacy of this resuscitation strategy in the respect of outcome of patients. Mtthod A total of 30 patients with septic shock were treated with the EDGT fluid treatment protocol as the conventional treatment group from December 2006 to June 2008; and another 26 patients were given fluid treatment under the guidance of PiCCO as PiCCO group from July 2008 to October 2009. Exclusion criteria included patients with history of heart and lung diseases, and liver and kidney dysfunction. The mean arterial blood pressure was maintained above or equal to 65mmHg in PiCCO group,and fluid resuscitation was concluded when global end-diastolic volume index reached 600～750 mL·-2 with the stroke volume variation in ＜ 10% and without auricular fibrillation. Nor-epinephrine was administrated to adiust the systemic peripheral vascular resistance index during 1300～1500 d·s·cm-5·m-2.Dobutamine was empoyed when global ejection fraction was compromised. The options of liquid and diuretics depend upon the presence d extra-vascular lung water. Central venous oxygen saturation and the level of lactate were observed 6hours after resuscitation. The liquid equilibrium for 3 days and the dosage of vasopressors were also recorded. The rate of survival, the time taken for weaning from mechanical ventilation, the days of ICU stay and rate of intact organ function within 28 days were compared between two groups. Results The demphics of patients of two goups were similar. There were no significant difference between PiCCO and the conventional group in values of central venous oxygen saturation and lactate 6 hours after admission to ICU (P ＞ 0.05). And 6 h and 1d after fluid resuscitation, the dosages of dobutamine and the dosages of nor-epinephrine used in PiCCO group were significantly higher than those in conventional group in which the dosages of dobutamine in two groups were [(145.4±24.5)mg vs. (104.2 ± 46.3) mg and (330.2 ± 30.3) mg vs. (202.4 ± 40.3) mg], respectively, and the dosages of nor-epinephrine [(14.5±3.8) mgvs. (10.2±5.6) mgand (38.2±4.2) rng vs.(20.1±6.2) mg], respectively. However, the dosages of vasopressors were similar between two groups 2 d and 3 d later. The amounts of liquid administered to get equilibrium in 6 h and the 3 d in PiCCO group were significantly less than those in conventional group [(2121±578) mL vs. (2910±987) mL and (3845±435) mL vs. (4545 ± 765) mL and (2467±510) mLvs. (2867±618) mL and (951±332) mLvs. (1472±533) mL], respectively. The days required to get free from mechanical ventilation within 28d were significantly longer in PiCGO group[(19.7 ± 8.3) d vs.(15.1±9.1) d], but the days of ICU stay were significantly shorter in PiCCO group [(7.5 ± 3.5) d vs. (9.5±3.2) d] (P＜0.05). The rates of survival and days free from organ failure within 28 days of two groups were similar(P＞0.05). Conclusions When the early fluid management guided with PiCCO in septic shock patients,the fluid management can be implement more safely and precisely. It can shorten the days of ICU stay and days of mechanical ventilation support with avoidance of fluid resuscitation.