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.

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 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.

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.

OBJECTIVETo investigate the expression of phospho-platelet derived growth factor receptor alpha (P-PDGFR-alpha) in gastrointestinal stromal tumors (GIST) and extra-gastrointestinal stromal tumors (EGIST) and its clinical significance.

METHODSExpression of P-PDGFR-alpha in 28 samples of positive CD117 and 13 samples of negative CD117 was detected by Envision immunohistochemical staining. Direct PCR sequencing was used to investigate the mutation status of c-kit gene exons 9, 11, 13, 17 and PDGFR-alpha gene exons 12 and 18.

RESULTSThe positive rate of P-PDGFR-alpha expression in GISTs with negative CD117 was 69.2%, which was significantly higher than that in GISTs with positive CD117 (7.1%, P<0.05). The positive rates of P-PDGFR-alpha expression in epithelioid GISTs(27.3%) and mixed GIST(63.3%) were both significantly higher than that in fusiform GISTs (9%, P<0.05). The positive rate of CD117 expression in fusiform GISTs (53.6%)was significantly higher than that in epithelioid GISTs (7.1%) and mixed GISTs(39.3%, P<0.05). C-kit gene mutation was found in 19 GIST cases with positive CD117. C-kit gene mutation was found in 19 of 28 GIST patients with positive CD117, among them, mutation of exon 11 occurred in 15 cases and exon 13 in 4 cases. No C-kit gene mutation was seen in 13 GIST patients with negative CD117. PDGFR-alpha gene mutation was found in 4 of 11 GIST cases with positive P-PDGFR-alpha and all occurred in exon 18.

CONCLUSIONSExamination of P-PDGFR-alpha expression may provide reliable evidence for the further improvement of pathological diagnosis,pathological typing and treatment for GISTs with negative CD117. Phosphorylated protein induced by PDGFR-alpha mutation may be associated with the important alternative molecular mechanism and the biological behavior of GIST development.

Adolescent ; Adult ; Aged ; Exons ; Female ; Gastrointestinal Stromal Tumors ; genetics ; pathology ; Gastrointestinal Tract ; pathology ; Humans ; Male ; Middle Aged ; Mutation ; Oxidative Phosphorylation ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Receptor, Platelet-Derived Growth Factor beta ; genetics ; metabolism ; Young Adult

A rapid analytical method was developed for the determination of N-methylcarbamoyl adduct in Hemoglobin of workers exposed to N,N-dimethylformamide by ultra high performance liquid chromatography-mass spectrometry ( UPLC/MS/MS). About 0. 1 g of hemoglobin sample, 40 μmol/L of 3-methyl-5-isopropylhydantoin (MIH) as the internal standard and 4. 75 mL of HCl-acetic acid (2∶1, V/V) were added in the centrifuge tube, and mixed for 3 min. Then the tube was heated in boiling water bath for 1h. After cooling down, 200 μL of the mixture and 600 μL of formic acid-acetonitrile (1%) were added into 96-well extract plate. The vacuum pump pressure was controlled to make the sample collection elute within 2-4 min.The purified collection was transferred into the sample vial, and 3-methyl-5-isopropylhydantoin ( MVH ) as degration product of N-methylcarbamoyl adduct was quantified by UPLC/MS/MS in multiple reaction monitoring ( MRM ) by internal standard method. A good linear relationship was obtained in the MVH concentration range of 0 . 01-1 . 0 μmol/L with the correlation coefficient of 0 . 999 . The recovery of added MVH in the blank sample was 97 . 3% and the relative standard deviation was 1 . 7%. The limit of detection (LOD) was 0. 01 μmol/g. This method was proved to be fast and efficient.