Objective To set up bronchodilation curve in patients with chronic obstructive pulmonary disease(COPD)or asthma and investigate the features of large and small airways dilation.Methods Pulmonary functions of 44 COPD and 68 asthma patients were determined before and after inhaling Formoterol.Results The curves of FEV1 and FVC rose at 15 minutes after inhaling Formoterol and then declined at 120 minutes post-formoterol in patients with COPD.The curves of FEV1/FVC and FEF75 presented as a wave form,and the curves of FEF50 and FEF75/25 rose at 30 minutes and declined after 30 minutes.The fitting quadr-curves model equations for FEV1,FVC and FEF50 over time were statistically significant(P

AIM: To construct the shuttle plasmid vector for thymidine kinase (tk) and EGFP fusion protein gene driven by IGF - Ⅱ P3 promoter, and investigate the specific killing effect of the HSV - tk/GCV system on hepatocellular carcinoma(HCC) cells in vitro. METHODS: Recombinant shuttle plasmid vector was constructed by techniques of genetic recombination and screening, and identified by restriction digestion and sequencing analysis. Then the recombinant shuttle plasmid was transfected into HepG2 and HeLa cells by techniques of lipofectamine transfection and its expression was detected by fluorescence microscope and RT -PCR. Cell killing after ganciclovir(GCV) application was determined by MTT. RESULTS: Identification of pDC316 -tkEGFP- P3 by enzyme digestion and sequencing analysis showed that the length, inserted location and direction of the target genes which were inserted into the recombinant were correct. It was found that enhanced green fluorescence protein could only be seen in HepG2 cells, but not in HeLa cells. The results of RT -PCR showed that only two bands could be seen in the samples of pDC316 -tkEGFP- P3 transfected HepG2 cells. The MTT test showed the selective cytotoxicity of GCV to the transfected HepG2 cells. CONCLUSION: The shuttle plasmid vector carrying the tkEGFP fusion protein gene driven by IGF - Ⅱ P3 promoter has been constructed successfully and its specific expression in HepG2 cells provided a sound basis for targeted gene therapy for HCC.

Objective To analyze the risk factors of pulmonary embolism in patients with negative Ddimer in serum in order to determine the need of pulmonary computed tomography angiograph (CTA) to confirm the final diagnosis in those patients for avoidance of misdiagnosis.Methods A retrospective analysis of 106 patients suspected to suffer from pulmonary embolism (PE) with serum negative D-dimer checked with pulmonary CTA was carried out.According to the results of CTA, the patients were divided into two groups, namely PE group (n =41) and non-PE group (n =65).The difference in clinic presentation, the time elapsed from onset to visit, N-terminal pro-brain natriuretic peptide (NT-proBNP), high risk factors (such as immobilization for 3 weeks, leg swelling and pain to palpation, history of deep vein thrombosis, malignancy) and Wells score (≥ 4 points indicates probability of PE).And logistic regression analysis was made to investigate the risk factors in PE with negative D-dimer.Results The analysis study showed that 38.6％ of total patients suspected to suffer from PE with serum negative D-dimer were checked by CTA to confirm the presence of PE.One important characteristics of the D-dimer negative PE patients was the longer time consumed from onset to visit [(9.51 ±2.01) d vs.(4.01 ±1.92) d, P＜ 0.05], and majority of the CTA positive patients suspected to suffer from PE with negative D-dimer had high risks of PE (P ＜0.01).Compared with the non-PE group, the Wells score ≥4 points and the level of serum NT-proBNP significantly increased in the PE group (P ＜ 0.01).Logistic regression analysis revealed that dyspnea, high NT-proBNP level and Wells sore ≥ 4 points were risk factors for D-dimer negative PE.Conclusion Delayed treatment was the main cause of misdiagnosis of D-dimer negative PE.Dyspnea, high NT-proBNP level and Wells sore ≥4 points were risk factors for suspected PE patients with negative D-dimer, and these patients should be confirmed by pulmonary CTA.On the contrary, PE could be excluded if patients with D-dimer negative had no these risk factors.

AIM: To investigate the genetic and epigenetic alterations of p14~ ARF gene and mutation status of p53 gene in human primary colorectal carcinomas and to analyze the relationship between the two gene changes and the role of abrogation of the p14~ ARF -p53 pathway in colorectal carcinogenesis. METHODS: The homozygous deletions, mutations, methylation of 5′ CpG islands, mRNA expression of p14~ ARF gene and mutations of p53 gene were assessed by PCR, direct sequencing, methylation-specific PCR, and RT-PCR in the tumorous and matched adjacent normal colorectal tissues from 56 patients with colorectal carcinoma. RESULTS: ① p14~ ARF alterations were detected in 27% (15/56) of colorectal carcinoma tissues studied, of which 1 case showed homozygous deletion, 14 cases showed 5′ CpG island methylation, and no mutation was found in any tumor. ②15 colorectal carcinomas with p14~ ARF alterations indicated lack of (13 cases) or at low level of expression (2 cases) of p14~ ARF mRNA, while expression of the p14~ ARF transcript was detected in the remaining 41 colorectal carcinomas and any matched adjacent normal colorectal tissues. ③ The mutations of p53 gene were detected in 48% (27/56) of colorectal carcinomas investigated. ④ Of these 56 cases, 12 had p14~ ARF alterations alone, 24 had p53 mutations alone, 3 had both p53 mutations and p14~ ARF methylation, and 17 had neither. 70% (39/56) of the samples had either or both abnormalities of the two genes, and p14~ ARF hypermethylation was related to wildtype p53 (P

Objective To evaluate the value of NT-proBNP in predicting weaning outcomes of the patients with mechanical ventilation by using the receiver operating characteristic (ROC)curve. Method The data of patients after the weaning of mechanical ventilation and spontaneous breathing trial (SBT) in the intensive care unit,from July 2008 to January 2010, were retrospective reviewed. All patients were divided into the success group and failure group as per the outcomes of weaning. Demographics and the serum NT-proBNP levels measured before weaning were compared between two groups with Student t -test and Chi-square test. The ROC curve was drawn to evaluate the value of serum NT-proBNP in predicting outcomes after weaning. Results A total of 160 patients were eligible for inclusion in the study, and there were 106 cases in success group and 54 cases in failure group.Compared with the failure group, the patients of success group were younger (63.17 ± 17.00 vs. 71.28 ± 12.56,t = 2.063,P =0.024), and no difference in gender (χ2 = 0.06, P ＞ 0.05). The NT-proBNP levels of failure group were significantly higher than those of success group (Lg NT-proBNP 2.80 ± 0.72 vs. 3.75 ± 0.56, t =2.351,P =0.014). The area under cure (AUC) of the ROC cure of NT-proBNPto predict the failure of weaning was 0.855 ±0.036 (95%CI0.784 ～ 0.925) when the cut-off level of NT-proBNP was 3635.5 pg/mL. And, this NT-proBNP level had a followed predictive efficiency in weaning outcome (Youden's index: 0. 60, accuracy:82.5%, sensitivity: 75%, specificity: 84.7%, positive likelihood ratio: 4.90, negative likelihood ratio:0.295,Kappa value: 0.62). Conclusions The levels of NT-proBNP before weaning have predictive value in weaning outcome, and it may be used as one of the screening indicators for weaning.

Objective To evaluate (1,3)-β-D-glucan (BG) assay as an aid for invasive fungal infection (IFI) diagnosis in severe pneumonia patients (diagnosis followed 2007 American Thoracic Society (ATS) and Infectious Disease Society of America (IDSA) severe pneumonia standard).Methods BG antigenemia was measured by BG Assay Box.IFIs was classified according to the blood fungal laboratory reports.Results 558 patients (185 females,373 males,mean age 64.7) were included.41 patients were proven to be fungal infected to be classified in exposure group.BG assay mean value in exposure group and unexposure group were (568.53 ±796.57) pg/mL,(51.4 ±63.27) pg/mL,respectively.Patients in the exposure group had significantly higher BG assay value than patients in the unexposure group (P ＜0.05).For the cutoff 100 pg/mL recommended by manufacturer,the sensitivity,specificity,positive predict value and negative predict value of the BG assay were 92.7％,92.5％,49.4％ and 0.6％,respectively.Conclusion BG assay has positive clinical value in invasive fungal infection diagnosis in severe pneumonia patients.

ObjectiveTo investigate whether esmolol could improve clinical outcome and tissue oxygen metabolism by controlling heart rate (HR) in patients with septic shock.Methods A single-center double-blinded randomized controlled trial was conducted. The patients suffering from septic shock received 6-hour early goal directed therapy (EGDT) with pulmonary artery wedge pressure≥ 12 mmHg (1 mmHg = 0.133 kPa) or central venous pressure (CVP)≥ 12 mmHg requiring norepinephrine to maintain mean arterial pressure (MAP)≥ 65 mmHg and HR≥95 bpm admitted to intensive care unit (ICU) of Guangdong General Hospital from September 2013 to September 2014 were enrolled. They were randomly divided into esmolol group and control group by computer-based random number generator. All patients received conventional basic treatment, while those in the esmolol group received in addition persistent esmolol infusion by micro pump with dosage of 0.05 mg·kg-1·min-1 with the dosage adjusted to maintain HR lower than 100 bpm within 24 hours. The patients in control group did not receive drug intervention for HR. The primary end-points consisted of length of stay in ICU and 28-day mortality. The secondary end-points included hemodynamic parameters [HR, MAP, CVP, cardiac index (CI), stroke volume index (SVI), systemic vascular resistance index (SVRI)] and tissue oxygen metabolism parameters [central venous oxygen saturation (ScvO2), lactate level (Lac)]before and 24, 48, 72 hours after the treatment.Results A total of 48 patients with septic shock were enrolled with 24 patients in esmolol group and 24 in control group.① The primary end-points: compared with control group, the length of stay in the ICU in the esmolol group was significantly shortened (days: 13.75±8.68 vs. 21.70±6.06,t = 3.680, P = 0.001), and 28-day mortality was significantly lowered [25.0% (6/24) vs. 62.5% (15/24),χ2 = 6.857,P = 0.009].② The secondary end-points: there were no significant difference in the hemodynamic and tissue metabolism parameters before treatment between two groups. No significant difference was found between before and after treatment of all above parameters in control group. HR and Lac in the esmolol group were obviously declined, SVI, SVRI, ScvO2 were gradually increased, but no significant difference in MAP, CVP, and CI was found. Compared with the control group, HR in the esomolol group was significantly lowered (bpm: 84.4±3.5 vs. 111.2±7.2,P< 0.01), SVRI and ScvO2 were significantly increased from 24 hours [SVRI (kPa·s·L-1·m-2): 137.9±1.6 vs. 126.9±1.3, ScvO2: 0.652±0.017 vs. 0.620±0.017, bothP< 0.01]; SVI was significantly increased (mL/m2: 39.9±2.2 vs. 36.8±1.7,P< 0.01) and Lac level significantly declined from 48 hours (mmol/L: 2.8±0.3 vs. 3.4±0.3,P< 0.01).Conclusion The results demonstrate that HR controlled by a titrated esmolol infusion given to septic shock patients was associated with an improvement in tissue metabolism, reduction in the length of ICU stay and lowering of 28-day mortality.

Objective To investigate the clinical value of the ratio of plasma vascular endothelial growth factor level to platelet count (VEGF/PLT) in predicting 28-day prognosis in patients with sepsis.Methods A prospective cohort study was conducted.From September 2009 to March 2013,164 sepsis patients in Intensive Care Unit (ICU) of Guangdong General Hospital were included for study.Patients with age younger than 18 years old,the illness already reaching final stage of chronic diseases,suffering from two or more organs dysfunction within 3 days,acute pancreatitis without infection,or less than 28 days of expected survival time were excluded.Finally,135 patients were included in the further analysis.Peripheral blood samples were collected at admission.Routine blood tests were done,and then VEGF levels in plasma were measured by enzyme linked immunosorbent assay (ELISA).Acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) scores were recorded every day for 7 days.Patients' prognosis was assessed during the following 28 days.The patients were divided into 28-day survival group and non-survival group.Comparison between two groups was done by single factor analysis.Spearman rank correlation was used to analyze the correlation between VEGF levels and PLT.Mutivariate logistic regression analysis was performed to identify the independent risk factor for 28-day prognosis.Receiver operating characteristic curve (ROC curve) was plotted,and the effect of related indexes on predicting 28-day survival was evaluated by area under ROC curve (AUC).Results There were no significant differences in VEGF (ng/L:471.73 ± 198.34 vs.383.49 ± 266.54,t=-1.918,P=0.057),PLT (× 109/L:220.40±127.60 vs.246.42± 100.72,t=1.275,P=0.204),leucocyte counts (× 109/L:12.48 ±4.62 vs.13.70 ±5.97,t=1.063,P=0.292),mean arterial pressure [mmHg (1 mmHg=0.133 kPa):86.50 ± 12.04 vs.91.03 t 13.10,t=1.557,P=0.123] and blood lactic acid (mmol/L:1.79 ± 1.30 vs.1.50 ± 0.60,t=-1.768,P=0.079) at admission between the non-survival group (n=42) and survival group (n=93).VEGF/PLT (2.59 ± 1.44 vs.1.73 ± 1.13,t=-3.756,P=0.000) as well as APACHE Ⅱ scores (15.50 ± 4.50 vs.13.28 ± 4.61,t =-2.022,P=0.045) of the non-survival group were significantly higher than those of survival group,and oxygenation index (PaO2/FiO2) of the non-survival group was significantly lower than that of survival group (kPa:32.38 ± 11.12 vs.37.04 ± 10.97,t=2.278,P=0.024).Correlation analysis showed that the concentration of VEGF was positively correlated with PLT (r=0.271,P=0.001).It was shown by multivariate logistic regression analysis that only VEGF/PLT was the independent risk factor in predicting 28-day prognosis in patients with sepsis [odds ratio (OR) was 1.591,95％ confidence interval (95％CI) 1.164-2.175,P=0.004].AUC of VEGF/PLT was 0.704 ± 0.047 (P=0.000,95％CI:0.611-0.797) for predicting 28-day survival.The optimal cut-off point was 1.32,and the sensitivity and specificity were 81.0％ and 48.4％,respectively.Conclusion VEGF/PLT can be used as one of the indicators to predict 28-day survival in patients with sepsis.

Objective To analyze the potential factors facilitating post-pyloric placement of spiral naso-jejunum tube in critically ill patients.Methods A retrospective study was carried out in patients requiring enteral nutrition (EN) from Apr 2005 through Dec 2011 in Intensive Care Unit (ICU).Severity of illness was assessed with APACHE Ⅱ score (acute physiology and chronic health evaluation Ⅱ).A selfpropelled spiral naso-jejunum tube was placed and observed for 24 hours.The forward movement and place of the tube tip was checked by bedside X-ray.The APACHE Ⅱ score,therapeutic measures,agents administered within 24 hours after tube insertion were recorded.The patients were divided into the success group and the failure group identified by bedside X-ray whether the tube tip entered into jejunum or not.Univariate analysis and multivariate Logistic regression analysis were used to find out the potential factors impacting on the success or failure in post-pyloric placement of naso-jejunum tube.Results A total of 508 patients composed of 337 male and 171 female,and aged (62.0 ± 19.2) years with APACHE Ⅱ score of (21.9 ± 7.3) were enrolled for study.The placement was successful in 205 (40.4％) of 508 patients.Univariate analysis showed that APACHE Ⅱ score ≥ 20,sedatives and analgesics,catecholamines,prokinetics,artificial airway and mechanical ventilation were potential factors facilitating the post-pyloric placement of naso-jejunum tube.Multivariate logistic regression identified that APACHE Ⅱ score ≥ 20,sedatives and analgesics and prokinetics were independent factors facilitating the post-pyloric placement of naso-jejunum tube.Conclusions The success rate of self-propelled spiral nasojejunal tubes insertion was relatively low.The prokinetics contributed higher success rate of naso-jejunum tube placement than factors of APACHE Ⅱ score ≥ 20,sedative and analgesic,catecholamine drugs,artificial airway and mechanical ventilation.There were no effects of age and gender on the placement of naso-jejunum tube.

Objective To explore whether hypertonic saline would partake in regulating Notch signaling in microglia in experimentally induced cerebral ischemic rats.Methods Male SD rats were randomly divided into sham group, cerebral ischemia group, normal saline group ( NS group ) , 10%hypertonic saline group (10%HS group) , the model of cerebral ischemia were established in all rats except the sham group by using middle cerebral artery occlusion ( MCAO) .After 2 hours of MCAO, the rats were through reperfusion for 24 h.In addition, rats in the normal saline group and 10% HS group were respectively treated with a continuous intravenous injection of normal saline (0.3 mL/h) and 10%HS (0.3 mL/h) by tail vein for 24 h.Immunofluorescence methods, RT-PCR and Western blot were used to detect the expression of Notch1 and intracellular Notch receptor domain ( NICD) .All data was analyzed by one-way analysis of variance ( ANOVA) , The intergroup comparisons were analyzed by the least-significant-difference (LSD) tests.Differences were considered statistically significant if P<0.05.Results Immunofluorescence showed that the expression of Notch1 and NICD were significantly increased in the microglia around peri-ischemia area in cerebral ischemia group and normal saline group compared to sham group;the expression of Notch1 and NICD in the microglia around peri-ischemia area were significantly reduced in 10% HS group compared to ischemia group and NS group.RT-PCR showed that the mRNA expression of Notch1 was significantly increased in ischemia group and NS group compared to sham group ( sham group: 1.000 ± 0.076; ischemia group: 2.203 ±0.283; NS group: 1.616 ±0.185; P <0.01 ); however, it was significantly reduced in 10% HS group compared to ischemia group and NS group ( ischemia group:2.203 ±0.283; NS group: 1.616 ±0.185; 10%HS group: 1.202 ±0.177; P <0.05 ) .Western blot showed that the protein expression of Notch1 was significantly increased in ischemia group and NS group compared to sham group ( sham group: 0.290 ±0.079; ischemia group: 0.750 ±0.029; NS group:0.765 ±0.182;P<0.01);but was significantly reduced in 10%HS group compared to ischemia group and NS group ( ischemia group:0.750 ±0.029; NS group:0.765 ±0.182;10%HS group:0.390 ±0.195;P<0.05 ) .The protein expression of NICD was significantly increased in ischemia group and NS group compared to sham group ( sham group: 0.401 ±0.196; ischemia group: 0.906 ±0.359; NS group:0.847 ±0.153;P<0.01);but was significantly reduced in 10%HS group compared to ischemia group and NS group ( ischemia group:0.906 ±0.359; NS group:0.847 ±0.153;10%HS group:0.561 ±0.165;P<0.05 ) .Conclusion Our results suggest that HS markedly suppresses Notch signaling in microglia around the ischemia tissue area in experimental induced cerebral ischemic rats.