Objective To investigate dynamic changes of vascular endothelial growth factor(VEGF)and endostatin(ES)in guinea.Pig of asthma and its effect on airway remodeling.Methods 40 guinea-pigs were randomly divided into normal control group(C),NS group(N),3d asthma group(A1),7d asthma group(A2)and 21d asthma group(A3),8 pigs ofeach group·Nothing was done on C Group guinea.Pigs.N group were intraperitoneal injected 2ml NS on the Ist day and the 8th day.NS inhalation started on the 15th day,1 time every day,3-5rain for each times,lasted for 3 day.Al,A2 and A3 Group were injected with 10rag ovalbumin(OVA),which made of aluminum hydroxide 20mg suspension in place of2ml NS,in place ofNS inhalation for3d,7d and 21d.At the end of24h-shot line after anesthesia BAL.Enzynle-linked immunosorbent(ELISA)was used to determine the concentration of VEGF and ES in bronchoalveolar lavnge fluid collection(BALF).Image analysis was used to determine airway circumference(Pi),outer circumference(Pe),and calculate the wall area(WA),airway smooth muscle area(SMC-A),airway smooth muscle cells(N).WA,SMC-A and N were standardized by Pi.Results With the extension of the duration of asthmatic guinea.Pigs,WA/Pi,SMC-A/Pi,N/Pi and concentration of VEGF in the airway were increased.Compared with group C and N,SMC-A/Pi,N/Pi and concentration of VEGF in asthma group were siguitlcanfly different.WA/Pi were siguificanfly different in A2 group and A3 group.The relation of the concentration of VEGF in Asthma guinea-pigs airway and WA/Pi was positive correlation.VEGF/ES and WA/Pi was highly positive correlated.Conclusions VEGF/ES inlbalance may play all important role in the airway remodeling of asthmatic guinea-pigs.

A rapid, sensitive and convenient LC-MS/MS method was developed for the determination of γ-aminobutyric acid (GABA) in human plasma. d2-γ-Aminobutyric acid (d2-GABA) was synthesized as internal standard (IS). After extraction from human plasma by protein precipitation with acetonitrile, all analytes were separated on a Luna HILIC column (100 mm x 3.0 mm, 3 μm) using an isocratic mobile phase of water: acetonitrile: formic acid (20 : 80 : 0.12) with a flow rate of 0.5 mL x min(-1). Acquisition of mass spectrometric data was performed in multiple reaction monitoring mode (MRM) in positive electrospray ionization using the transitions of m/z 104 --> 69 for GABA and m/z 106 --> 71 for d2-GABA. The method was linear in the concentration range of 5.00 to 1 000 ng x mL(-1). The intra- and inter-day precisions were within 9.9%, and accuracy ranged from 99.1% to 104%, within the acceptable limit across all concentrations. The method was successfully applied to a pharmacokinetic study of GABA tablets in healthy Chinese volunteers.

A simple and rapid method was developed based on high performance liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) to determine sivelestat and its metabolite XW-IMP-A in human plasma. After a simple protein precipitation, the samples and internal standards were analyzed on a C18 column by a gradient elution program. The mobile phase consisted of 30% acetonitrile in methanol and 5 mmol · L(-1) ammonium acetate at a flow rate of 0.7 mL · min(-1). The mass spectrometric data was collected in multiple reaction monitoring mode (MRM) in the negative electrospray ionization. The standard curves were linear in the range of 10.0-15,000 ng · mL(-1) for sivelestat, and 2.50-1000 ng · mL(-1) for XW-IMP-A. The low limits of quantitation were identified at 10.0 and 2.50 ng · mL for sivelestat and XW-IMP-A, respectively. The intra- and inter-day precision were within 11.3% and 13.1% for sivelestat and XW-IMP-A, and accuracy was 0.3% and 0.6% for sivelestat and XW-IMP-A, within the acceptable limits across all concentrations. The method was successfully validated in the pharmacokinetic study of sivelestat in healthy Chinese volunteers.

Objective To observe the effects of transdermal acupoint electric stimulation(TEAS)assisting sodium nitroprus-side induced controlled hypotension on serum glucose (Glu),angiotensin Ⅱ (ATⅡ)and superoxide dismutase (SOD),and to inves-tigate the protective effect of TEAS under controlled hypotension anesthesia.Methods 60 cases undergoing elective endoscopy si-nus surgery by adopting sodium nitroprusside induced controlled hypotension under general anaesthesia maintained the mean arterial pressure(MAP)in 50-60 mm Hg and were randomly and equally divided into two groups.The group Ⅰ conducted TEAS,while the group Ⅱ did not conduct TEAS.The controlled hypotension time and surgery time were recorded in the two groups;Glu and ATⅡ values were detected before anesthesia (T0 ),30 min after hypotension (T1 ),hypotension stopping(T2 );SOD was detected at T0 ,T2 ,30 min after hypotension(T3 ).Results The operation time and controlled hypotension continuous time had no statistically significant difference between the two groups(P >0.05).The Glu value in the group Ⅰ had no statistically significant difference a-mong the 3 time points,while which at T1 ,T2 was higher than that at T0 in the group Ⅱ(P <0.05),and which at T1 ,T2 in the group Ⅱ was higher than that in the group Ⅰ (P <0.05);the ATⅡ value at T1 was higher than that at T0 in the group Ⅰ (P <0.05),while which at T1 ,T2 was higher than that at T0 in the group Ⅱ(P >0.05),which at T1 ,T2 in the group Ⅱwas higher than that in the group Ⅰ (P <0.05);the SOD value at T2 was lower than that at T0 in the group Ⅰ,which at T2 ,T3 in the group Ⅱwas lower than that in the group Ⅰ(P <0.05).Conclusion TEAS assisting sodium nitroprusside controlled hypotension can better in-hibit the stress response.

A simple, sensitive, selective, and reproducible liquid chromatography-tandem mass spectrometric method was developed for the simultaneous determination of repaglinide and metformin in human plasma using d5-repaglinide and d6-metformin as internal standards (ISs). After a simple protein precipitation using acetonitrile as the precipitation solvent, both analytes and ISs were separated on a Venusil ASB C 18 (150 mm x 4.6 mm, 5 microm) via gradient elution using acetonitrile--10 mmol x L(-1) ammonium acetate as the mobile phase. A chromatographic total run time of 7.5 min was achieved. Mass spectrometric detection was conducted with atmospheric pressure chemical ionization under positive-ion and multiple-reaction monitoring modes. The method was linear over the 0.2 to 60.0 ng x mL(-1) concentration range for repaglinide and over the 4 to 1 000 ng x mL(-1) range for metformin. For both analytes, the intra- and inter-accuracies and precisions were within the +/- 15% acceptable limit across all concentrations. The validated method was successfully applied to a clinical bioequivalence study.

ObjectiveTo evaluate the primary effect of granulocyte-monocyte colony stimulating factor (GM-CSF) as an immunotherapy option for treatment of residual disease after alloHSCT.Methods Immunotherapy was performed on two patients with blood malignancy to treat residual disease after allo-HSCT. The patient one,who was diagnosed as having MDS-RAEB Ⅱ,showed bone marrow displasis and incomplete chimerism 6 months after unrelated donor HSCT.Immunosuppressive drug was withdrawn without induction of graft-versus-host disease (GVHD).The patient two B-ALL demonstrated a residual disease at molecular level 30 days post-transplantation.Both of them were given GMCSF (300 μg) subcutaneously once every two days for totally three weeks.During the whole period,skin itch and rash,liver function,subgroups of lymphocytes,and MDSCs and DCs in peripheral blood were investigated.Results In case one,grade Ⅰskin acute GVHD (aGVHD) appeared as early as one week after GM-CSF administration,as well as grade Ⅱ (skin and liver) by the end of the third weeks,and GM-CSF injection was withdrawn.One month later since the start of GM-CSF,the patient showed normal bone marrow morphology and full donor type chimerism. Cyclosporine A (CsA), mycophenolate mofetil and methylprednisolone were administered for two weeks to control GVHD.In the other case,grade Ⅰ aGVHD occurred 9 days after GMCSF administration,and whole blood CsA maintained at 0.134-0.472 μmol/L.Prednisone (30mg per day for 5 days) was used to control grade Ⅱ GVHD from the 11th day after GM-CSF,and grade Ⅰ GVHD continued without any intervention.On the 30th day after GM-CSF treatment,bone marrow aspiration showed complete molecular remission.In both of the two cases,no differences in lymphocytic subtypes were revealed before and after GM-CSF administration,while there were trends of increased DC number and decreased MDSCs in peripheral blood.ConclusionThe administration of GM-CSF as an immunotherapy option for blood malignancy may contribute to the clearance of residual disease after Allo-HSCT.

Objective To observe the effects of ( - )-epigallocatechin-3-gallate (EGCG) on human prostate cancer xenografted tumor growth and connexin43 expression in nude mice,and explore the mechanism of the EGCG on prevention for prostate cancer.Methods The methyl thiazolyl tetrazolium and annexin-V/PI double-labeled flow cytometry methods were used to observe the growth inhibiting rate (IR)and apoptosis rate (AR) of human prostate cancer cell line PC-3 which was treated by EGCG at different concentration (10,20 and 40 mg/L,respectively).The scrape-loading fluorescence dye transfer method was applied to assess the gap junction intercellular communication (GJIC) through fluorescence microscope.PC-3 cells were subcutaneously transplanted to establish tumor-bearing nude mice model.A total of 32 mice were randomly divided into four groups,both control group and three treatment groups were treated with different doses of EGCG ( 10,20 and 40 mg/kg,respectively).After two weeks,the mice prostate tumor tissues were taken out.The tumor wet weight was measured and tumor growth inhibiting rate was calculated.The tumor microvascular density (MVD) and apoptosis index (AI) were detected by the immunohistochemical techniques and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling techniques,respectively.Semi-quantitative reverse transcription polymerase chain reaction was used to examine the expression level of the Cx43 mRNA.Results EGCG at concentration ( 10 and 20 mg/L) could significantly inhibit the proliferation[(22.33 ±4.62)％,(38.67 ±5.67)％ vs (3.47 ±0.31 )％,P ＜0.01],induce the apoptosis [(7.84 ± 1.37 ) ％,( 24.53 ± 2.28 ) ％ vs ( 2.17 ± 0.70 ) ％,P ＜ 0.01] and enhance the GJIC of PC-3 cells.EGCG of different doses could inhibit prostate cancer xenografted tumor growth,induce tumor cells apoptosis and inhibit angiogenesis.EGCG ( 20 and 40 mg/kg) could effectively up-regulate Cx43 mRNA expression in xenografted tumor (0.58 ± 0.08,0.80 ± 0.07 vs 0.42 ± 0.04,P ＜ 0.0 ).The effects had significant correlation with the dose-dependent of EGCG ( P ＜ 0.05 ).Conclusions EGCG could up-regulate the Cx43 expression and enhance the gap junction intercellular communication mediated by Cx43 in the prostate tumor,which provide the experimental evidence for the mechanism of its effectively inhibiting the prostate cancer growth.

Objective To explore the clinical value of ureteroscopy and minimally invasive percutaneous nephrolithotomy ( MPCNL) in the treatment of ureteral calculi patients with acute renal dysfunction .Methods Clini-cal data of 124 ureteral calculi patients with acute renal dysfunction were retrospectively analyzed .86 cases were trea-ted with holmium-laser lithotripsy under ureteroscope .38 cases were treated with MPCNL under the guide of ultra-sound.Results Three months after operation ,the stone clearance rate was 100%,and no severe complications were observed.The renal function decreased to normal levels in 102 cases(82.3%).Conclusion The holmium laser lith-otripsy under ureteroscope and MPCNL can deal with double sites of ureteral calculi ,and offer advantages with less in-vasion,safety and efficiency ,which can be the first choice for the ureteral calculi combined with acute renal dysfunc -tion.

OBJECTIVETo investigate the effects of lung protective ventilation strategy combined with lung recruitment maneuver on ARDS complicating patients with severe burn.

METHODSClinical data of 15 severely burned patients with ARDS admitted to our burn ICU from September 2011 to September 2013 and conforming to the study criteria were analyzed. Right after the diagnosis of acute lung injury/ARDS, patients received mechanical ventilation with lung protective ventilation strategy. When the oxygenation index (OI) was below or equal to 200 mmHg (1 mmHg = 0. 133 kPa), lung recruitment maneuver was performed combining incremental positive end-expiratory pressure. When OI was above 200 mmHg, lung recruitment maneuver was stopped and ventilation with lung protective ventilation strategy was continued. When OI was above 300 mmHg, mechanical ventilation was stopped. Before combining lung recruitment maneuver, 24 h after combining lung recruitment maneuver, and at the end of combining lung recruitment maneuver, variables of blood gas analysis (pH, PaO2, and PaCO2) were obtained by blood gas analyzer, and the OI values were calculated; hemodynamic parameters including heart rate, mean arterial pressure (MAP), central venous pressure (CVP) of all patients and the cardiac output (CO), extravascular lung water index (EVLWI) of 4 patients who received pulse contour cardiac output (PiCCO) monitoring were monitored. Treatment measures and outcome of patients were recorded. Data were processed with analysis of variance of repeated measurement of a single group and LSD test.

RESULTS(1) Before combining lung recruitment maneuver, 24 h after combining lung recruitment maneuver, and at the end of combining lung recruitment maneuver, the levels of PaO2 and OI of patients were respectively (77 ± 8), (113 ± 5), (142 ± 6) mmHg, and (128 ± 12), (188 ± 8), (237 ± 10) mmHg. As a whole, levels of PaO2 and OI changed significantly at different time points (with F values respectively 860. 96 and 842. 09, P values below 0. 01); levels of pH and PaCO2 showed no obvious changes (with F values respectively 0.35 and 3.13, P values above 0.05). (2) Levels of heart rate, MAP, CVP of all patients and CO of 4 patients who received PiCCO monitoring showed no significant changes at different time points (with F values from 0. 13 to 4. 26, P values above 0.05). Before combining lung recruitment maneuver, 24 h after combining lung recruitment maneuver, and at the end of combining lung recruitment maneuver, the EVLWI values of 4 patients who received PiCCO monitoring were respectively (13.5 ± 1.3), (10.2 ± 1.0), (7.0 ± 0.8) mL/kg ( F =117.00, P <0.01). (3) The patients received mechanical ventilation at 2 to 72 h after burn, lasting for 14-32 (21 ± 13) d. At post injury day 3-14 (7 ± 5) d, lung recruitment maneuver was applied for 2-5 (3.0 ± 2.0) d. All 15 patients recovered without other complications.

CONCLUSIONSLung protective ventilation strategy combining lung recruitment maneuver can significantly improve the oxygenation in patients with severe burn complicated with ARDS and may therefore improve the prognosis.

Acute Lung Injury ; physiopathology ; therapy ; Blood Gas Analysis ; Burns ; complications ; Extravascular Lung Water ; Hemodynamics ; Humans ; Positive-Pressure Respiration ; Respiration, Artificial ; methods ; Respiratory Distress Syndrome, Adult ; complications ; physiopathology ; therapy ; Treatment Outcome

Objective To analyze the motivation of Chinese patients with chronic myelogenous leukemia (CML) who have stopped the tyrosine kinase inhibitor (TKI). Methods Forty-seven CML patients who have stopped TKI provided informed consent prior to their participation in the study. These patients were divided into relapse and non-relapse group at the endpoint of the observation. None of the patients received any CML-associated therapies after TKI cessation. The reasons of withdrawal were analyzed statistically. Results The reasons for cessation included patient's request due to cost(59.57 %, 28/47), patient's plan to getting pregnant(8.52 %,4/47),side-effect of TKI(23.40 %,11/47)and other reasons(8.52 %,4/47).At the endpoint of observation, 23 patients suffered molecular relapse. Among them, 15 cases (65.22 %) were due to cost; 1 case (4.35 %) was due to getting pregnant, 5 cases (21.74 %) were due to side-effect and 2 cases (8.69 %) were due to other reasons. There was more frequency relapse in the group of insufficient cost. Conclusion The motivation of Chinese CML patients who have stopped TKI might show impact on the outcome,and the motivation is mainly related with history of drug reduction and withdrawal.