Objective To investigate the influence of propofol pretreatment on the increased glomerular endothelial cell permeability induced by lipopolysaccharide (LPS) in rats.Methods Glomerular endothelial cells isolated from SD rats were cultured in 24-well plates(200 μl/well) and transwell filters (100 μl/filter) at 1×106/ml and assigned into 6 groups (n=10 each):control group (group C) , introlipid group (group I), propofol group (group P) , LPS group (group L), LPS+introlipid group (group L+I) and LPS+propofol group (group L +P). In group I, 10% introlipid 4 μg/ml was added. In group P, 4 μg/ml propofol was added. In group L, 10 μg/ml LPS was added. In group L+I, 10% introlipid 4 fig/ml combined with 10 μg/ml LPS was added. In group L+ P, 4 μg/ml propofol combined with LPS 10 μg/ml was added. Introlipid or propofol was added 30 min before the administration of LPS and the corresponding concentrations mentioned above were all final concentrations.After 6 h incubation with LPS, the cells were collected for measurement of vascular endothelial growth factor (VEGF) mRNA expression using RT-PCR. The supernatant was collected for determination of the VEGF concentration by ELJSA. The endothelial cell permeability was determined. Results Compared with group C, the expression of VEGF mRNA was up-regulated and the VEGF concentration and endothelial cell permeability were significantly increased in L, L+I and L + P groups (P＜0.05 ) ,but no significant change was found in the parameters mentioned above in I and P groups (P＞0.05). Compared with group L, the expression of VEGF mRNA was downregulated and the VEGF concentration and endothelial cell permeability were significantly decreased in L+P group (P＜0.05), but no significant change was found in the parameters mentioned above in group L+I(P＞0.05). A positive correlation existed between the concentration of VEGF and the permeability of endothelial cells(r= 0.833,P＜0.05).Conclusion Propofol pretreatment can decrease the increased glomerular endothelial cell permeability induced by LPS probably through down-regulation of VEGF expression.

Objective To investigate the influence of propofol on endotoxin-induced expression of vascular endothelial growth factor (VEGF) receptor 2 in renal glomerular endothelial cells of rats.Methods The primarily cultured renal glomerular endothelial cells of Sprague-Dawley rats were seeded in 24-well plates at a density of 1×106 cells/ml (200 μl/well) and divided into 6 groups (n =35 each) using a random number table:control group (group C);introlipid group (group I);propofol group (group P);lipopolysaccharide (LPS) group (group L);LPS + introlipid group (group L+I);LPS + propofol group (group L+P).In group I,10％ introlipid was added with the final concentration of 4 μg/ml.In group P,propofol was added with the final concentration of 4 μg/ml.In group L,LPS was added with the final concentration of 10 μ g/ml.In group L+I,10％ introlipid was added with the final concentration of 4 μg/ml at 30 min before LPS with the final concentration of 10 μg/ml was added.In group L+P,propofol was added with the final concentration of 4 μg/ml at 30 min before LPS with the final concentration of 10 μg/ml was added.After 6 h of incutation,the cells were collected for measurement of cell permeability and VEGF receptor 2 mRNA expression (using RT-PCR),VEGF receptor 2 protein expression (by Western blot),and for examination of the morphology of cytoskeletal protein filamentous-actin (F-actin) with confocal microscope (by immunofluorescence).Results Compared with group C,the expression of VEGF receptor 2 mRNA and protein was significantly up-regulated,and the cell permeability was increased in L,L+I and L+P groups,and no significant changes were found in the parameters mentioned above in I and P groups.Compared with group L,the expression of VEGF receptor 2 mRNA and protein was significantly down-regulated,and the cell permeability was decreased in L+P group,and no significant changes were found in L+I group.F-actin connected closely between adjacent cells and a dense peripheral F-actin band was formed in C,I and P groups,while F-actin depolymerized,the peripheral F-actin band was disrupted,and cells shrank in L and L+I groups.In group L+P,a tighter intercellular connection of F-actin was observed and cytoskeleton was found to be intact.Conclusion Propofol can inhibit endotoxin-induced increase in the permeability of renal glomerular endothelial cells through down-regulating the expression of VEGF receptor 2 in rats.

Objective To explore the diagnostic value of gram-stained sputum smears in elderly patients with ventilator-associated lower respiratory infection (VALRI).Methods 368 elderly patients undergoing mechanical ventilation over 48 hours in ICU were retrospectiely analyzed from Jun 2009 to Jun 2011.42 patients fulfilled the inclusion criteria (with good-quality sputa and presence of the same microorganism in sputum and blood cultures).58 gram-stained smears and sputum cultures were obtained and analyzed.Results The diagnostic sensitivity and specificity of Gram-positive bacteria and Gram-negative bacteria by gram-stained smears were 82.35％ and 78.05％,80.49％ and 82.35％,respectively.The positive and negative predictive value of Gram-positive bacteria and Gramnegative bacteria were 63.64％ and 91.43％,91.67％ and 60.87％,respectively.The coincidence rates in diagnosing Gram-positive bacteria and Gram-negative bacteria were 81.03％ and 79.31％,respectively.Conclusions Gram-stained sputum smear is reliable in diagnosing ventilator-associated lower respiratory infection in elderly patients,and has a certain clinical significance in guiding to the selection of antibiotics.

Objective To investigate the incidence of brain injury and analyze the risk factors in very low birth weight infants (VLBWI),in order to explore methods to improve neurodevelopmental outcome.Methods This was a retrospective study in which we reviewed the incidence of brain injury in 326 VLBWI who admitted to our NICU from October 2007 to September 2014.The differences of general characteristics,high-risk perinatal factors,complications of prematurity and respiratory support in brain injury group (n =132) and no brain injury group(n =194) were analyzed,and Logistic regression was used to analyze the related risk factors.Results In 326 VLBWI,premature brain injury was observed in 132 cases(incidence of 40.49％).Hemorrhagic brain injury was observed in 80 cases(incidence of 24.54％,80/326),in whom total 66 cases (20.25％,66/326) with periventricular-intraventricular hemorrhage (PVH-IVH),41 cases with PVH-IVH grade Ⅰ-Ⅱ,25 cases with PVH-IVH grade Ⅲ-ⅣV.Non-hemorrhagic brain injury was observed in 52 cases (incidence of 15.95 ％,52/326),in whom 46 cases with periventdcular leukomalacia (14.11％,46/326).There were no statistically significant differences in general characteristics except gender differences between the two groups.Further analysis of the 28 factors including high-risk perinatal factors,premature complications and respiratory support between the two groups,showed that there were significant differences in 16 factors including asphyxia,chorioamnionitis,antenatal steroid use,neonatal respiratory distress syndrome,hypoxemia,apnea,pulmonary hemorrhage,patent ductus artedosus,hypotension,acidosis,hypocapnia,anemia,hyperglycemia,thrombocytopenia,high frequency ventilation and extrautedne growth retardation.Further binary Logistic regression analysis showed that antenatal steroids use was negatively correlated with brain injury,chorioamnionitis,acidosis,and patent ductus arteriosus were positively correlated with brain damage,the relative risks of premature brain injury was 0.108,7.222,3.045 and 4.565 respectively.Conclusion The mortality rate and the incidence of brain injury in VLBWI are still high.Chorioamnionitis,acidosis and patent ductus arteriosus are closely associated with premature brain damage.Antenatal steroids can prevent premature brain injury.We should pay much attention to these risk factors in the perinatal management of VLBWI,in order to decrease the incidence of premature brain injury.

To observe the expressions of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in rats with pulmonary fibrosis (PF) induced by bleomycin, and to explore the mechanisms of Shenlong Decoction in preventing and treating PF.

Objective To explore the effects of application of probiotics on intestinal flora and related cytokines in infants with cholestatic liver disease.Methods (1) Eighty-four infants with cholestatic liver disease had been hospitalized from October 2010 to June 2011 in the First Hospital of Guangxi Medical University.The infants with cholestatic liver disease were randomly divided into the probiotic intervention group and the non-probiotic intervention group.Quantification of intestinal bacteria was detected by SYBR Green Ⅰ real-time fluorescent quantitative polymerase chain reaction,then the number of 3 kinds of bacteria before and after the treatment was compared.(2) The indices of liver function,blood ammonia,cholesterol were detected.The levels of serum transforming growth factor-β1,tumor necrosis factor-α(TNF-α)and interleukin-6 (IL-6) were measured by enzyme linked immunosorbent assay.Results (1) The number of bifidobacterium,lactobacillus and bifidobacterium/escherichia coli (B/E) were significantly increased (all P ＜ 0.01),whereas the number of escherichia coli was significantly decreased in the probiotic intervention group(P ＜ 0.05),however,there were no differences in the non-probiotic intervention group(all P ＞ 0.05).(2)The indices of total billirubin,direct billirubin,γ-Glutamyltransferase,total bile acid,alanine aminotransferase,blood ammonia,alkaline phosphatase were significantly improved after therapy in 2 groups (all P ＜ 0.05).The levels of TNF-α and IL-6 were significantly decreased in the probiotic intervention group (t--7.31,P =0.00;t =-2.90,P =0.01),but there were no differences in the non-probiotic intervention group.The level of BA was significantly decreased in the probiotic intervention group than the non probiotic intervention group (t =-8.37,P =0.00).(3) The B/E value were significantly inverse correlated with level of serum IL-6 (r =-0.796,P =0.01).Conclusions It may help to restore the intestinal flora and balance the immune function in infants with cholestatic liver disease after application of probiotics.

ObjectiveTo investigate the therapeutic effect of human umbilical cord mesenchymal stem cell-paracrine substance on fulminant hepatic failure (FHF) rat, and to study the effect on liver function and hepatocyte proliferation. MethodsMesenchymal stem cells(MSCs)were separated from human umbilical cord, and surface makers of cells were detected by flow cytometry. Human umbilical cord mesenchymal stem cells-conditioned medium(MSC-CM) was prepared. FHF rat model was induced by intraperitoneal injection of D-galactosamine and they were randomly diveded into three groups: MSC-CM group, NS group, PHGF group. 24 h later, 1 ml MSC-CM, 1 ml 0. 9% NaCl solution and lml PHGF solution was injected into the tail vein of MSC-CM, NS, and PHGF rats, respectively. In each group (n=8 per group), blood samples were collected at 12, 24, 36, and 60 h after treatment from inner canthus for analysis of blood ALT and TBIL levels. We used five rats per group for tissue collection after sacrifice at 36 h after treatment and 10 animals per group for survival analysis. PCNA immunohistochemical staining was used in the sections of liver tissue to detect hepatocyte proliferation. Results24 h after treatment, the levels of ALT and TBIL in the MSC-CM and PHGF groups were lower than those in the NS group(P＜0. 05), but there was no significant difference between the MSC-CM and PHGF groups. There were more PCNA-positive hepatocytes in the MSC-CM and PHGF groups than in the NS group(P＜0.01), but there was no significant difference between MSC-CM and PHGF group. Survival analysis found that the survival rate of rats in the MSC-CM and PHGF groups was higher than that of rats in the NS group (P=0. 049), but there was no significant difference between the MSC-CM and PHGF group. ConclusionsThe paracrine substance of human umbilical cord mesenchymal stem cells can stimulate hepatocyte proliferation and improve liver function of FHF rats, potentially creating a new avenue for the treatment of FHF.

Objective To establish the long-term culture system for fetal skin fibroblast by performing long time in vitro cultivation of the cells,and study the potential of its differentiation to hepatocytes.Methods Fibroblast was isolated from human fetus skin tissue.Surface phenotypes of cells were detected by ICC and FCM,and biological characteristics were analyzed by the karyotype analysis and soft agar colony formation observation.ALB、CK18、CK19 were detected by ICC,glycogen stain by PAS,AFP and ALB mRNA by RT-PCR after P3～30cells were induced differentiation by cytokines of HGF,FGF4 and OSM.Results CD29,CD49f,HLA- Ⅰ and CD 105 were highly expressed while CD90 hardly in skin fibroblast.The rate of induced differentiation of fibroblast into hepatocyte-like cells was approximately 5％.The cells could be cultured in vitro for almost 50 passages with normal karyotype and no oncogenic and immortalized characteristics.Conclsion The skin fibroblast possesses the characteristic of mesenchymal stem cell and can be induced into hepatocyte-like cell in vitro.

Objective To explore the diagnostic value of sputum smear coupled with simplified clinical pulmonary infection score (CPIS) of ventilator associated pneumonia (VAP) in the early stage.Methods A cohort of 59 consecutive patients with VAP admitted in Intensive Care Unit from June,2014 to June,2016 were enrolled for a prospective and observational study.Concurrently,another 59 patients without pulmonary infection undergone mechanical ventilation over 48 hrs,were assigned into the control group.The criteria of exclusion were patients with pulmonary malignancies,autoimmune diseases and immunodeficiency.APACHE Ⅱ scores of all patients were recorded.All patients' inferior airway sputum which met the criteria was taken to make a validated sputum smear (i.e.polymorphonuclear leukocyte ＞ 25and squamous epithelial cell ＜ 10 per low-power field) for Gram stain and culture on the admission day.Meanwhile,simplified CPIS were calculated.Data were statistically processed by SPSS 15.0,enumeration data were statistically analyzed by Chi-Square test,and measurement data were represented as Mean ± SD.The significant differences in characteristics between two groups were analyzed by independent t test,and P ＜ 0.05 was considered statistically significant.As positive sputum smear and simplified CPIS ≥ 5 were set respectively as a positive screening criterion,sensitivity,specificity,positive predictive value and negative predictive value of each marker and combined markers were calculated.Results There were no significant differences in demographics and clinical features (including age,sex,APACHE Ⅱ scores) of patients in VAP and non-VAP patients (P ＞ 0.05).The rates of bacteria detected were Gram-negative [44.1％ (26/59)],Gram-positive [40.6％ (24/59)],none [10.2％ (6/59)] and both [5.1％ (3/59)] bacteria in VAP group,while [39.0％ (23/59)],[30.5％ (18/59)],[27.1％ (16/59)] and [3.4％ (2/59)]were found in non-VAP group correspondingly.There were no significant differences in the percentages of different bacteria in sputum smear between two groups (P ＞ 0.05).The values of diagnostic sensitivity of sputum smear and sputum smear coupled with simplified CPIS were 89.8％ and 84.7％;the specificity were 27.1％ and 79.7％;the positive predictive values were 55.2％ and 80.6％;and the negative predictive values were 72.7％ and 83.9％,respectively.Conclusions No matter the ventilated patients suffered VAP or not,bacteria might be detected from their lower respiratory tracts.Sputum smear could not be taken as an exclusively diagnostic evidence.While sputum smear coupled with simplified CPIS might improve the diagnostic efficacy of VAP,and provide the guildlines of appropriate choice of antibiotics employed in the early stage.

Objective:To evaluate the role of silencing information regulator 1 (SIRT1)/nuclear factors E2-related factor2 (Nrf2) signaling pathway in berberine-induced reduction of renal ischemia-reperfusion (I/R) injury in mice.Methods:Thirty SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 18-22 g, were divided into 5 groups ( n=6 each) using a random number table method: sham operation group (S group), renal I/R group (RIR group), berberine+ I/R group (B group), berberine+ I/R+ SIRT1 inhibitor EX527 group (BE group) and berberine+ I/R+ Nrf2 inhibitor ATRA group (BA group). After the right kidney was removed, the left renal artery was clamped for 45 min followed by reperfusion to establish the model of renal I/R injury.In B, BE, and BA groups, berberine 100 mg·kg -1·d -1 was given for intragastric administration at 14 days before surgery.In group BE and group BA, EX527 5 mg·kg -1·d -1 and ATRA 10 mg·kg -1·d -1 were injected intraperitoneally at 3 days before surgery, respectively.The equal volume of normal saline was given for 14 consecutive days in group S and group RIR.Blood samples were collected from orbital vein at 24 h of reperfusion for measurement of serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations, for determination of the interleukin-1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) contents (by enzyme-linked immunosorbent assay) and expression of SIRT1, Nrf2, apoptosis-associated speck-like protein containing CARD (ASC), caspase-1, nucleotide-binding oligomerization domain-like receptor containing pyrin domain (NLRP3) (by Western blot) and for examination of the pathological changes of renal tubules (with a light microscope). The damage to the renal tubules was scored. Results:Compared with group S, the concentrations of serum Cr and BUN, the contents of renal IL-1β and TNF-α and renal tubular injury score were significantly increased in RIR, B, BE and BA groups, the expression of SIRT1, Nrf2, ASC, caspase-1 and NLRP3 was up-regulated in RIR, BE and BA groups, and the expression of SIRT1, Nrf2, caspase-1 and NLRP3 was up-regulated in group B ( P<0.05). Compared with group RIR, the concentrations of serum Cr and BUN, the contents of renal IL-1β and TNF-α and renal tubular injury score were significantly decreased in B, BE and BA groups, the expression of SIRT1 and Nrf2 in group B, Nrf2 and ASC in BE group and SIRT1, ASC and caspase-1 in BA group was up-regulated, and the expression of ASC, caspase-1 and NLRP3 in group B, SIRT1 and NLRP3 in BE group and Nrf2 in BA group was down-regulated ( P<0.05). Compared with group B, the serum concentrations of Cr and BUN, the contents of IL-1β and TNF-α and renal tubular injury score were significantly increased in BE and BA groups, the expression of ASC, caspase-1 and NLRP3 in BE and BA groups was up-regulated, and the expression of SIRT1 in BE and Nrf2 in BA groups was down-regulated ( P<0.05). Conclusion:SIRT1/Nrf2 signaling pathway is involved in the process of berberine-induced reduction of renal I/R, which is related to inhibiting pyroptosis in mice.