AIM: To validate the alternations of flow velocity patterns in the infarct-related artery (IRA) during no-reflow phenomenon in a canine model of acute myocardial ischemia and reperfusion by transthoracic Doppler echocardiography (TTDE) combined with myocardial contrast echocardiography (MCE) by means of administration of Albunex. METHODS: Nineteen dogs first underwent 60 min myocardial ischemia and then followed by 60 min,120 min and 180 min reperfusion ( n = 6, 6 and 7, respectively). The perfusion defect area determined by MCE at 60 min myocardial ischemia was regarded as risk area (RAMCE). The perfusion defect area defined by MCE after reperfusion was considered as no-reflow area (NRAMCE). The ratio between NRAMCE and RAMCE ≥ 25 %was regarded as the development of no-reflow phenomenon and the ratio of NRAMCE to RAMCE＜25% was considered as the myocardial reflow. The coronary flow velocity parameters in IRA were determined through TTDE. RESULTS: Two dogs died during experiment and the remaining seventeen dogs completed throughout the procedure.There were seven dogs in reflow group and ten dogs in noreflow group. No significant difference was present in reflow group between at baseline and at 60 min reperfusion in systolic peak velocity (PVs), systolic velocity time integral (VT Is), corrected systolic flow duration (cFDs),diastolic peak velocity (PVd), diastolic velocity time integral (VT Id), corrected diastolic flow duration (cFDd),diastolic deceleration rate (DDR), corrected diastolic deceleration duration (cDDD) (P＞0.05), however, a significant difference was found in no-reflow group between at baseline and at 60 min reperfusion in PVs,VTIs, cFDs, PVd, VTId and cFDd (P＜0.05). The most marked alterations during diastolic phase were the increase of DDR and reduction of cDDD. CONCLUSION: The impaired microvasculature may profoundly affect the coronary flow velocity pattern in the IRA. The increase in microvascular resistance and decrease in coronary perfused pressure can contribute to the changes.Transthoracic Doppler echocardiography combined with MCE has the capability of noninvasive assessment of coronary flow velocity pattern in the IRA during no-reflow phenomenon.

Objective The purpose of this study was to investigate the role of p38 mitogen-activated protein kinase ( p38 MAPK) in the lung injury induced by mechanical ventilation.Methods Fifteen healthy 80 day-old pigs weighing (22.5　? 1.5)kg were randomly divided into three groups according to the tidal volume(VT) and PEEP of mechanical ventilation: group A (VT = 16ml?kg-1, PEEP = 0) ; group B (VT = 6 ml?kg-1, PEEP= 16cm H2O) and group C(VT = 16ml?kg-1, PEEP = 8cm H2O). The animals were mechanically ventilated for 3h, then sacrificed by exsanguination. Right lower lobe was immediately removed for identification of intercellular adhesion molecule-1 ( ICAM-1 ) expression using immunohistological technique, determination of phosphorylated p38 MAPK content using Western Blot and microscopic examination. Results There was significant histological changes in the lung tissue in group A and B, but no significant histological changes were found in group C. The expression of ICAM-1 was positive in the lung in group A and B but negative in group C. The level of phosphorylated p38 MAPK among the 3 groups. Conclusion Acute lung injury can be induced by mechanical ventilation with high tidal volume or low tidal volume plus high PEEP, p38 MAPK may mediate the inflammatory response-induced lung injury.

OBJECTIVE:To evaluate the quality of TCM decoction pieces in our hospital preliminarily and explore the solu-tions for the problem of the quality of decoction pieces purchased by the hospital. METHODS:In Jun. 2012,a system was estab-lished by our hospital,where the quality inspector was designated to daily inspect new batches of TCM decoction pieces such as ap-pearance provided by suppliers. The batches of TCM decoction piece samples inspected and those of unqualified products from Jun. 2012 to May 2014 were calculated. The reasons of unqualified products were analyzed and corresponding solutions were made. RE-SULTS&CONCLUSIONS:Over two years,a total of 94 671 batches of TCM decoction piece samples were inspected by our hos-pital,among which 737 were unqualified products predominantly because of mildew,damage by worms,greasing,containing for-eign substances and others. The solutions to such problems included interviewing the suppliers,returning or discontinuing the use of crude drugs and special focus on particular seasons and on the demand for key varieties. The unqualified products rate in the quality inspection reduced from 0.56%in Jun. 2012 to 0.34%in May 2014. Therefore,setting the post of drug quality inspector in the hos-pital can ensure the quality of TCM decoction piece purchased,however,it need to improve the inspection and acceptance of quali-ty inspection by sampling,scoring and using the new technology.

Objective To explore the risk factors causing tetany in patients with hyperventilation syndrome. Meth?ods A total of 103 patients with hyperventilation syndrome treated in our hospital were included in this study. According to whether there was tetany, patients were divided into tetany group and non-tetany group. Values of gender, age, electrolyte, pH and p(CO2) were analysed between two groups. The factors of P<0.1 were engaged in binary Logistic regression. Logistic regression (Forward Wald) was used to analyze the risk factors of tetany in patients with hyperventilation syndrome. Re?sults In 103 patients there were 70 patients with tetany (68%), 33 patients without tetany(32%). The serum K+, serum phos?phorus and p(CO2) values were significantly lower in tetany group than those of non-tetany group (P<0.01), while the pH value was significantly higher in tetany group than those of non-tetany group (P<0.01). There were no significant differences in gen?der, age, serum Na+, serum Cl-, serum calcium (bound calcium and ionized calcium), ionized calcium and serum Mg2+levels be?tween two groups (P>0.05). It was revealed that the younger age, the lower level of the serum K+, serum phosphorus and p(CO2) were the risk factors of tetany through binary Logistic regression analysis. Conclusion The risk factors of tetany in patients with hyperventilation syndrome include younger age, lower level of serum K+and serum phosphorus and reduced p(CO2).

Aim: To study the effects of molecule weight,the N/P ratio,solvent and ionic strength on the formation and surface properties of PEI/DNA complexes,and study its transfection efficiency.Methods: The N/P ratio of the prepared PEI/DNA complexes was optimized using gel electrophoresis and UV quantitative assay.The particle size and surface charge of the complexes were measured in different solvents and ionic strength.The transfection effi-ciency in HepG2 cells was observed.Results: There was a positive correlation between combination of PEI and DNA and molecule weight of PEI.In addition,the surface properties of PEI/DNA complexes were also influenced by the solvents and ionic strength.Comparable transfection efficiencies in HepG2 cells were observed for the Lipo-fectamine 2000/DNA and the prepared PEI(25 kD)/DNA complexes in PBS at the N/P ratio of 12-15,which was much higher than that of naked DNA.Conclusion: The optimized PEI/DNA complexes could effeciently transfect the cells in comparison to the positive control.

Objective To investigate the role of Xuebijing injection in inhibiting perioperative inflammatory responses and protecting the function of multiple organs.Methods A single-blind,randomized,parallel controlled trial was conducted.60 patients in the First Affiliated Hospital of Zhejiang University School of Medicine,aged 18 to 80 years,ASA grade Ⅰ-Ⅲ,undergoing elective abdominal surgery,were enrolled.The patients were randomly divided into the control group (n =30) and the treatment group (n =30).In the control group,after induction of anesthesia,a continuous infusion of 0.9％ normal saline (NS) 200 mL was given in a speed of 2 mL/min,while a continuous infusion of Xuebijing 2 mL/kg in 100 mL of 0.9％ NS was given at 2 mL/min in the treatment group after induction of anesthesia.The blood sample was drawn,and body temperature,routine blood test,C-reactive protein (CRP),liver and kidney function,fasting glucose (Glu),and serum interleukin-6 (IL-6),high mobility group protein B 1 (HMGB 1) levels were determined in all the patients before anesthesia (T1),at the end of operation (T2),12 hours after operation (T3),or at 5:00 am on the third day after operation (T4).At the same time the adverse reactions were recorded for evaluation of the safety of Xuebijing.Results After using Xuebijing injection,T3 body temperature and the T3-T1 temperature difference in treatment group were significantly lower than those of the control group(℃℃:36.70 ± 0.37 vs.37.38 ± 0.47,t＝6.199,P＝0.000; 0.07 ± 0.50 vs.0.85 ±0.58,t＝5.598,P＝0.000).Postoperative white blood cell count,neutrophil percentage,and CRP were significantly higher than those before the operation,but the differences between two groups were not statistically significant.Compared with the control group,alanine aminotransferase (ALT),aspartate transaminase (AST),total bilirubin (TBil) levels at T3 of treatment group were significantly reduced [ALT (U/L):17.56 ± 9.80 vs.88.60 ± 179.76,AST(U/L):27.53 ± 13.12 vs.84.16 ± 151.14,TBil(μ,mol/L):15.46 ± 9.79 vs.25.63 ± 25.33,all P＜0.05].Difference of conjugated bilirubin (CB),blood urea nitrogen (BUN),creatinine (Cr),Glu were not statistically significant between two groups.IL-6 showed an increasing trend after the operation in both groups,and IL-6 level (ng/L) at T2 of the treatment group was significantly lower than that of the control group (41.42 ± 59.74 vs.124.84 ± 119.66,t=3.405,P=0.001).The HMGB 1 level of two groups at T4 were lower than those at T1,but it decreased significantly only in treatment group (μg/L:22.03 ± 15.73 vs.45.09 ± 33.79,P＜0.05),and there was no significant difference between two groups.No serious adverse events occurred during the clinical trial.Conclusions Application of Xuebijing injection during anesthesia can significantly diminish postoperative inflammatory injury,which plays an important role in the protection of liver function,helps restore organ function and improve prognosis,and it is safe and effective.

Objective To investigate the expressions of interferon γ(IFNγ), interferon inducible protein-10(IP-10). chemokine receptor CXCR3 and their significance in infection-associated hemophagocytic syndrome(IAHPS). Methods Forty-three patients with IAHPS, 35 infection patients without HPS and 25 healthy controls were included in the study. The serum IFNγ and IP-10 levels were measured by enzyme linked immunosorbent assay(ELISA). The expression of CXCR3 on cell surface of CD_4~+ and CD_8~+ T cells in peripheral blood was determined by flow cytometry. SPSS 13.0 was used for data processing, and independent-sample t test was performed to compare the differences among the groups. Results The serum IFNγ and IP-10 levels in patients with IAHPS were( 608±135) pmol/L and(939±141) pmol/L respectively, which were significantly higher than those in without HPS group[(154±45) pmol/L and (385±119) pmol/L, t=4.97 and 4.02, P<0.05] and healthy controls[(56±18) pmol/L and (248±98) pmol/L, t=5.27 and 4.77, P<0.05]. The expressions of CXCR3 on CD_4~+ and CD_8~+ T cells in IAHPS group were (35±11)% and (23±8)% respectively, which were significantly higher than those in without HPS group[(24±7)% and (16±7)%, t=3.12 and 3.62, P<0.05] and healthy controls[(20±6)% and (12±5)%, t=4.46 and 4.93, P<0.05]. Conclusion The expressions of IFNγ, IP-10 and CXCR3 are increased significantly in patients with IAHPS, which may be related to the disease pathogenesis.

Objective To investigate the correlation between gene polymorphism within human β defensin 1 (DEFB1) and fungal susceptibility to severe sepsis through case-control association study.Methods A total of211 patients with severe sepsis in ICU were enrolled in the present case control study. Sepsis in this study was diagnosed according to the definition of American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference in 1992 and 2002. Based on the development of fungal infection during ICU stay, all 211 patients were divided into fungal infection group (Group Ⅰ) and control group (Group C). Alleles and genotypes of-1816A/G, -390A/T, -52A/G, -44C/G and-20A/G within DEFB1 gene were assayed in all 211 patients by means of DNA direct sequencing, Allele-specific PCR amplifications or high-throughput site-specific TaqMan assay. Genetic analysis was employed to calculate the distribution frequency of haplotypes. The correlation between the genomic variations (allele,genotype and haplotype) and fungal infection was analyzed by Chi-square test or Fisher's exact test.Odds ratio (OR) was employed to reflect the correlation degree of genetic factor with fungal susceptibility to severe sepsis. Results Group Ⅰ enrolled 80 patients, of whom 43 pstients were male, at age of (60.81 ± 18.30) years. Group C enrolled 131 patients, of whom 80 patients were male, at mean age of (60.42 ± 17.03) years. No significant difference was found between two groups in aspect of gender and age (P＞0.05). The genetic locus of -1816A/G, -390A/T, -52A/G, -44C/G and -20A/G of both groups were in agreement with Hardy Weinberg equilibrium. No significant difference was found between two groups in the distribution of allelic frequencies and genotype frequencies (P ＞0.05). No significant difference was found in the distribution frequency of four common haplotypes of the above five genetic locus such as AAACG, ATGCA, GTGGG and ATACG (all P ＞ 0.05). Conclusions Genetic locus of -1816A/G, -390A/T, -52A/G, -44C/G and-20A/G within DEFB1 gene have no correction with fungal infections in severe sepsis, suggesting that DEFB1 gene polymorphism may not serve as a key genetic marker for the predisposition to fungal infection in severe sepsis.

Objeoctive To compare the effects of etomidate and propofol on cerebral oxygen metabolism in patients undergoing abdominal surgery.Methods Thirty-six ASA Ⅰ or Ⅱ patients aged 30-64 yr weighing 42-73 kg undergoing abdominal surgery under general anesthesia were randomly divided into 2 groups (n=18 each):group E etomidate and group P propofol.Left radial artery was cannulated for continuous direct BP monitoring.A catheter was inserted into right internal jugular vein(LJV) and advanced cephalad until jngatar bulb for blood sampling.Both groups received midazolam 0.08 ms/ks,fentanyl 3μg/kg and vecurunium 0.1 mg/kg and in addition group P received propofol 1.5 mg/ks and group E etomidate 0.3 mg/kg respectively for induction of anesthesia.Anesthesia was maintained with propofol infusion at 4-6 mg·kg-1·h-1 in group P and etomidate infusion at 0.4-0.7 mg·kg-1·h-1 in group E and intermittent iv boluses of fentanyl and vecuronium.ECG,MAP,HR,SpO2 and PET CO2 were continuously monitored.Blood samples were taken from radial artery and IJV for blood gas analysis and lactic acid measurement before induction of anesthesia(T1),immediately after intubation (T2),30 min after skin incision (T3) and at the end of operation(T4).The rate of cerebral O2 extraction (CERO2) was calculated.Results The hemodynamic variables were within the normal range throughout the anesthesia and operation.The oxygen saturation and oxygen partial pressure of both arterial and venous blood(SaO2,SjvO2,PaO2,PjvO2) rose significantly after induction of anesthesia in both groups.There was no significant difference in arterial and venous blood lactic acid level and SaO2,SjvO2,CaO2,CjvO2,Da-jvO2 or CERO2 at all time points between the two groups.Conclusion Both etomidate and propofol combined with midazolam and fentanyl can decrease cerebral O2 metabolic rate and there is no significant difference between the two groups.

When we try to establish the gene recombinant yeast cell to screen the androgenic endocrine disruptors, the key procedure is the androgen receptor (AR) expression in the yeast cell. For this purpose, we obtained the GPD (glyceraldehyde-3-phosphote dehydrogenase) promoter from the yeast genosome of W303-1A using PCR system and inserting it into Swa I and BamH I sites of pYestrp2. The new constructed vector was named pGPD. The V5 epitope tag DNA with a 5'-BamH I and a 3'-EcoR I sticky end was cloned into the corresponding site of the pGPD vector to yield the vector of pGPDV5. The 2 723 bp full length AR ORF amplified by PCR from pcDNA3.1/AR was fused to V5 epitope tag DNA in pGPDV5 to give the AR yeast expression vector of pGPDV5/AR. This fused vector was transformed into the yeast cell (W303-1A). Western blot was used to detect the V5 fused protein of AR, in the protocol of which the primary monoclonal antibody (IgG(2a)) of mouse anti-V5 and the polyclonal secondary antibody of goat anti-mouse (IgG) linked to horseradish peroxidase (HRP) were used to detect the specific protein in the given sample of the transformed yeast extract. The result showed that the fused protein of AR was expressed successfully in the yeast cell.
Base Sequence ; Endocrine Disruptors ; analysis ; Epitopes ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Glyceraldehyde-3-Phosphate Dehydrogenases ; genetics ; Humans ; Molecular Sequence Data ; Promoter Regions, Genetic ; Receptors, Androgen ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; genetics ; Yeasts ; genetics ; metabolism