Objective To evaluate the abilities and thresholds of stroke volume variation (SVV) and pleth variability index (PVI) in predicting fluid responsiveness during increased intra-abdominal pressure.Methods 28 patients undergoing laparoscopy-assisted radical gastrectomy were selected.PV1 was continuously displayed by the Masimo.Radical 7.All patients were also monitored with Vigileo/FloTrac system.Haemodynamic data such as MAP,HR,SVI,SVV,PI,PVI and C VP were recorded before and after volume expansion(HES 6％,7ml/kg).Fluid responsiveness was defined as an increase in SVI≥ 15％ (△ SVI ≥ 15).Results The SVV threshold of 9.5％ before volume expansion was able to diserimihate the responders from the non-responders with a sensitivity of 100％,and a specificity of 63.6％.The threshold for PVI was 14.0％,the sensitivity of 100％ and specificity of 81.8％ were obtained.There was no significant difference between the area under the receiver operating characteristics (ROC) curves of SVV and PVI(0.981,0.939,respectively),and there was significant correlation between the baseline SVV and the baseline PVI(r =0.740,P ＜ 0.01).Conclusion SVV and PVI can predict fluid responsiveness accurately during increased intra-abdominal pressure,the baseline SVV is correlated well with baseline PVI,and the ability of SVV and PVI in predicting fluid responsiveness is similar.

BACKGROUND:Hyperbaric oxygen as a wel-recognized therapy for ischemic and hypoxic diseases has to be combined with other treatments. OBJECTIVE:To study the effects of hyperbaric oxygen combined with neural stem cel transplantation via tail vein in chronic stress depression rats. METHODS:Of 60 Sprague-Dawley rats, 15 rats randomly selected were given no treatment as normal group, and the rest 45 ones were used to establish a rat model of depression and randomly divided into three groups:model group (n=15, without giving any treatment), neural stem cel group (n=15, injection of 1 mL neural stem cel suspension (3×106) via the tail vein) and combined group (n=15, hyperbaric oxygen treatment plus neural stem cel injection). Hyperbaric oxygen treatments were carried out four times per day, for 1 week, and al the treatments were given 24 hours after modeling. Rats in each group were subjected to body mass measurement and sugar water consumption test 1 and 2 weeks after treatment, and open field test within 5 minutes after treatment to observe rat behavior changes. The survival and distribution of CM-Dil labeled neural stem cel s were observed by fluorescence microscopy. The changes in the size, morphology and number of hippocampus neurons in rat were detected by the method of nylon staining. TUNEL method was used to measure the apoptosis of nerve cel s. RESULTS AND CONCLUSION:Compared with the normal group, in the model group, rat’s body weight, sucrose water preference and open field test scores were significantly lower, reduced number of hippocampal neurons with intact structure was found and TUNEL results showed more apoptosis (P<0.05). Compared with the model group, the body weight, sucrose preference and open field test score increased significantly (P<0.05), the number of hippocampal neurons was increased, while the number of apoptotic cel s was reduced significantly in the neural stem cel and combined groups (P<0.05). Compared with the neural stem cel group, these indexes were improved more significantly in the combined group (P<0.05). More CM-Dil positive cel s were found in the combined group than the neural stem cel group (P<0.05). Additional y, the number of hippocampal neurons was higher in the combined group than the neural stem cel group, but lower than the normal group. To conclude, transplantation of neural stem cel s combined with hyperbaric oxygen can improve the depression behavior and apoptosis in the hippocampal neurons in chronic stress depression rats.

Objective To search for a good method for generation of hematopoietic stem cells (HSC) by embryonic stem cells (ESC),and to investigate the potential of HSC derived from ESC to reconstruct hematopoiesis in vivo.Methods Using a three-step method to induce a mice ESC line, E14.1-into HSC.And identifying HSC by flow cytometry analysis cell markers with CD34 +/Sca-1+, then HSC (1×109L-1)from ESC differentiation were injected into severe combined immunodefieency (SCID) mice for observing teratoma formation.To validate function of HSC by colonngenic cells assay and to reconstitute the hematopoiesis in lethally irradiated mice.Results Combining to use more hematopoietic stimulating factor to availably promote the El4.1 cell into embryoid body (EBs) with abundant hematopoietic progenitors.EBs were induced after 14 day to fast differentiate for HSC with peak percentages of CD34+/Sca-1+ cells reached to (13.72±2.07)%.To harvested ceils from EBs by day 14 for second -step HSC differentiation, percent of CD34+/Sca-1+ cells rise to(24.62±2.50) % after day 16 induction.Cloning forming units (CFU) analysis showed that more Erythro -myeloid cloning generation was observed at this period.Cells obtained in the second step are subsequently plated on monolayer of mice bone marrow stromal cells, in the presence of TPO, FLt3 ligand and superoatant of mice fetal liver stromal cells, cultured for additional 15 days, followed fast expansion of CD34+/Sca-1+ to maximally (58.64±4.20 ) % with more CFU-E, CFU-GM and CFU-GEMM population.Wright-Giemsa stain showed that its had the character of hematopoietic progenitors.Cells from the third-step were injected into SCID mice, but no teratomas were recovered in 2 mices after 6 weeks.Positive selection of CD34+/Sca-1+ cells by magnetic sorting from the third - step differentiation were transplanted into 7 lethally irradiated female mice while predominant hematopoietic reconstitution were observed in 10 days after transplantation, with 71.4% successful engraftment rate.And 3 recipients showed that the cell population of the peripheral blood leukocytes,red cells, hemoglobin approached normal index at 40 days after transplant, hut followed relative'slow renew in platelet count.Survival rate of transplant group is 43.0%, compared to 100% mortality in control mice.Karyotyping assays confirmed female mice with XY.Conclusions The results showed that the three-step differentiation and the culture conditions described here good support differentiation of ESC into HSC.HSC derived from ESC were safe without teratomas formation in body, and its can reconstruct hematopoiesis.

Objective To investigate whether human umbilical vein endothelial cells as a feeder layer was capable of supporting the growth of embryonic stem cells in vitro.Methods Human umbilical vein endothelial cells were isolated and cultured and then prepared as feeder cells after 3 passages.Alkaline posphatase activity(AKP) staining,stem cell surface marker test and karyotypes were conducted in different periods of cell culture.The suspension of stem cells cultured after 20 passages on endothelial cells were inoculated to the legs of severe combined immunodeficiency(SCID) mice subcutabeously to test the teratoma formation.Results E14.1 embryonic stem cells retained good colonies when they were cultured on endothelial cells for 3 passages and 8 passages.In addition,they expressed SSEA-1,Oct-4,and a membrane alkaline phosphatase to a high extent at passages 3 and 8.Embryonic stem cells cultured for 15 passages stably retaind a normal karyotype.Embryonic stem cells cultured on endothelial cells for 20 passages were inoculated into the hind leg of SCID mouse,teratomas containing three embryonic layers were recovered six weeks later.Conclusion Human umbilical vein endothelial cells would support effectively embryonic stem cells expansion,and provide a clinically safe method to expand ES cells for futureclinical application.

AIM:To investigate the potential of hematopoietic stem cells(HSCs)derived from mouse embryonic stem cells(ESCs)to reconstruct hematopoiesis in vivo.METHODS:Using a three-step method,a mice embryonic stem cell line,E14.1 was induced into hematopoietic stem cells.The cell markers with CD34+/Sca-1+ were identified by flow cytometry analysis,then HSCs(1?109 cells/L)from third-step were injected into SCID mice for observing teratoma formation.To validate function of HSCs,colonogenic cell assay was conducted and the hematopoiesis in lethally irradiated mice was reconstituted.RESULTS:The method of three-step differentiation,combined to use more hematopoietic stimulating factor promoted the E14.1 cell differentiation into HSCs with highest percent of CD34+/Sca-1+ cells(as high as 58.64%?4.20%)with more CFU-E,CFU-GM and CFU-GEMM populations.The cells showed the character of hematopoietic progenitors by Wright-Giemsa staining.Positive selected CD34+/Sca-1+ cells by magnetic sorting from third-step differentiation were transplanted into 7 lethally irradiated female mice while predominant hematopoietic reconstitution were observed in 10 d after transplantation,with 71.4%(5/7)successful engraftment rate.Three recipients showed that the cell population of the peripheral blood leukocytes,red blood cells and hemoglobin approached to normal index at 40 d after transplantation,but followed relative slow renew in platelet count.Survival rate in transplant group was 43%,compared to 100% mortality in control mice.Karyotyping assays confirmed the female mice with XY.CONCLUSION:The three-step differentiation and the culture conditions described here support the differentiation of mouse ESCs into HSCs.HSCs derived from mouse ESCs can reconstruct hematopoiesis.

Implementation of information technology in clinical research has resulted in revolutionary changes in drug development. Based on the good clinical practice (GCP) requirements on data, processes and documentations, and the era of fast growth in clinical studies using up-to-date information technology, we explore an integrated e-clinical solution in clinical studies in China.
China ; Clinical Trials as Topic ; Data Collection ; methods ; Medical Informatics ; methods

Objective To compare the efficacy of laryngeal mask airway Supreme (LMAS) and Streamlined Liner of the pharynx Airway (SLIPA) in patients undergoing general anesthesia. Methods Eighty ASA Ⅰ or Ⅱ patients aged 18-70 yr weighing 45-80 kg undergoing general anesthesia were randomly divided into 2 groups ( n = 40 each): group LMAS and group SLIPA. Pharyngeal airway was inserted after induction of anesthesia with fentanyl 3μg/kg and propofol 2.0-2.5 mg/kg. MAP, HR, number of insertion, rate of successful placement at first attempt, placement time, airway sealing pressure, peak and mean airway pressure and side effects were recorded. Results There were no significant differences in MAP, HR and rate of successful placement at first attempt between the two groups. The placement time was significantly longer, the airway sealing pressure lower and the incidence of side effects higher in SLIPA group than in LMAS group. There was no significant difference in the peak and mean airway pressure between the 2 groups. Conclusion Both LMAS and SLIPA can assure good airway sealing and adequate ventilation. The complication is rare. The efficacy of LMAS is better.

Objective This study is to explore the differences in the curative effect of intra -abdominal infusion between Recombinant mutant human tumor necrosis factor and Dendritic cell -Cytokine induced killer . Methods We selected 48 advanced gastric cancer patients with ascites .Those patients were randomized into two groups:DC-CIK group and rmhTNF group .After one month treatment ,we observed the adverse events both two groups,and evaluated the clinical beneficial responses ,the responsive rate ,the tumor indicators′level,the immune indexes and the time of tumor progression .Results In the DC-CIK group,the RR and the CBR were 83.3%and 66.67%,respectively;while in the rmhTNF group,they were 58.33%and 75%,respectively.The difference between two groups was statistically significant (P<0.05).Only CA724 decreased after treatment in two groups (P=0.015).There were no significant differences of tumor markers between the two groups before and after treatment(P>0.05).The ratios of CD3 +、CD4 +、NK cells distinctly increased after treatment (P<0.05),but the ratio of CD8 +cell was no obvious difference in DC -CIK group(P=0.551);The ratio of NK cells increased obviously after treatment(P=0.027),but the ratios of CD3 +and CD4+were similar as previous in rmhTNF group(P>0.05).One year follow-up of the time to progression(TTP)was 7.1 months in DC-CIK group,and 5.8 months in TNF group,which was statistically significant (P=0.02).Conclusion Both rmhTNF and DC-CIK can improve the efficacy for the patients with malignant ascites caused by gastric cancer .However ,DC-CIK immunotherapy shows better active effect on improvement of specific immune function .

Objective To explore whether human umbilical venous endothelial cells could be used as feeder layer to support the growth of embryonic stem cells (ESC) and keep ESC undifferentiated.Methods The venous vessels of umbilical cord obtained from healthy puerperal were perfused with collagenase.The isolated endothelial cells went through primary culture and passages for expansion.Factor Ⅷ antigens determination was implemented.Endothelial cells with good growth and 3 or above passages were treated with mitomycin-C(10 mg/L) and prepared as feeder layer,on which E14.1 ESC was transplanted for subculturing to observe the morphological characterization and determine ESC alkaline psphatase (AKP) activity and the expression of stem cell marker Oct-4.Severe combined immune deficiency(SCID) mouse in vivo terotoma formation experiment was performed to identify its pluripotent properties.Results Human umbilical vein-derived endothelial cells grew well in culturing in vitro and regenerate in large numbers.The endothelial cells maintained normal cellular morphological and biological characterization after 10 passages.The cells stopped proliferating after being treated with mitomycin-C,but its activity and morphological properties were well-maintained with 24 hours,which was a fundamental property of serving as feeder layer.E14.1 ESC remained undifferentiated in human umbilical venous endothelial cells after 3-8 passages,the cells grew in colony and showed high expression of AKP and stem cell Oct-4.In vivo pluripotency experiment showed that 6 weeks after being transplanted to SCID mice E14.1 ESC of 6 and 10 passages in endothelial cells both could form teratoma containing 3 layers of tissue cells.Conclusions Human umbilical venous endothelial cell serve as a convenient feeder layer cell with rich sources.It can effectively support ESC growth and heterogenous and prevent the heterogeneous protein pollution and pathogenic microorganisms caused by animal cell feeder layers,thus solve the problem of biological safety of ESC clinical application.