Objective The study aimed to investigate the effect of lipoxin A4 (LXA4) on lipopolysaccharide (LPS)?induced oxidant stress in human renal tubular epithelial cells (HK2 cells) and possible underlying mecha?nisms. MethodsHK2 cells were divided into three groups: Control ,LPS and LPS+LXA4 groups. After cells were treated with indicated conditions,morphological changes were observed. The expressions of Nrf2 were detected by immunofluorescence and cells were collected for RT?PCR experiments.Results HK2 cells seemed disrupted and necrotic with the administration of LPS. However ,LXA4 could prevent cells from injury induced by LPS. LPS decreased Nrf2 expression and promoted it to translocate to cytoplasm ,while LXA4 could increase its expression and promote it to translocate to nucleus. Moreover ,LPS could decrease Nrf2 and its downstream molecule mRNA expressions,but LXA4 could reverse this effect. Conclusion Our results demonstrated that LXA4 effectively inhibit?ed HK2 cell oxidant stress via Nrf2 pathway.

Objective To investigate the dynamic changes of soluble urokinase-type plasminogen activator receptor (suPAR) and its predictive value in late-onset sepsis in the newborn.Method To collect the data of neonates aged 7 days and older,who were diagonsed to have infections.They were admitted to neonatal intensive care unit of our Hospital from January 2014 to January 2015.The group of sepsis and nonseptic group were assigned according to the diagnostic criteria of sepsis,and a control group was selected without infection.Blood cultures were collected in patients on the first day when infection was identified and the serum suPAR and CRP were measured on the first day,fourth day and tenth day respectively.The controls were tested with suPAR and CRP when infection was excluded.The levels of blood suPAR and CRP in the three groups were compared and the receiver-operating characteristic curve was performed according to serum suPAR level of neonates with sepsis on the first day.Result A total of 65 infants with infections (40 were septic and 25 were non-septic) were enrolled in this study and 20 patients were selected as control group.There were significant differences in serum suPAR and CRP levels between the patients with and without infection (P ＜ 0.001).The level of suPAR in the survivors of the sepsis group was significantly decreased as time went by,and the difference was statistically significant on the 10th day compared with the 1 st day [9.3 (8.2,13.1) ng/ml vs.18.9 (14.8,24.7) ng/ml,P ＜ 0.05].The level of CRP increased first initially and then decreased with time,while the highest level was on the 4th day and the difference was statistically significant compared with the 10th day [19.0 （ 6.8,56.4) mg/L vs.6.4 (2.5,12.0) mg/L,P ＜ 0.05].The levels of serum suPAR and CRP in non-sepsis group were not significantly different (P ＞ 0.05).There were no deaths in the sepsis group and the non-septic group,but the levels of suPAR between survivals and deaths in the infection groups were statistically significant [15.4(10.6,21.6) ng/ml vs.22.6 (15.4,31.9) ng/ml,Z =-2.063,P =0.039].The area under the receiver-operating characteristic curve of serum suPAR was 0.955 (95％ CI 0.906 ～ 1.000,P ＜0.001),and the sensitivity was 90％ and the specificity was 100％ when the suPAR level was 10.9 ng/ml.Conclusion Early elevated serum suPAR levels were prominently related to the severity of neonatal late-onset sepsis.The level of first day suPAR has a high sensitivity and specificity in the prognosis of sepsis and can be helpful to predict the prognosis.

Objective To research the mechanism of High Volume Hemofiltration (HVHF) on acute lung injury (ALI) induced by LPS in dogs. Methods After injection of LPS (650 ?g/kg) via central vein within 30 min, Sixteen healthy hybrid male dogs were divided into control group and treatment group randomly (n=8). PaO2、PaCO2 in artery blood were recorded. Contents of TNF-?、IL-6 and IL-10 in plasm were measured by radioimmunity. The activity of NF-?B in lung homogenate was measured by flow cytometer. The content of surfactant protein B (SP-B) in lung homogenate was measured by Western-blotting.Changes of lung histopathology was observed via electron microscopy. Results After injection of LPS, PaO2 and PaO2/FiO2 began to decrease. PaO2 and PaO2/FiO2 in treatment group kept higher than that in control group (P

Genetic transformation is an effective method to improve breeding objective traits of orchids. However, there is little information about genetic transformation of Cymbidium sinensis. Rhizomes from shoot-tip culture of C. sinensis cv. 'Qijianbaimo' were used to establish a practical transformation protocol of C. sinensis. Pre-culture time, concentration and treating methods of acetosyringone, concentration of infection bacteria fluid (OD600), infection time, and co-culture time had significant effects on β-glucuronidase (GUS) transient expression rate of C. sinensis cv. 'Qijianbaimo' rhizome. The GUS transient expression rate of rhizome was the highest (11.67%) when rhizomes pre-cultured for 39 d were soaked in bacterium suspension (OD600 = 0.9) supplemented with 200 μmol/L acetosyringone for 35 min, followed by culturing on co-culture medium supplemented with 200 μmol/L acetosyringone for 7 d. Under this transformation conditions, 3 transgenic plantlets, confirmed by GUS histochemical assay and PCR, were obtained from 400 regenerated plantlets, and the genetic transformation rate was 0.75%. This proved that it was feasible to create new cultivars by the use of Agrobacterium-mediated genetic transformation in C. sinense.
Agrobacterium ; Coculture Techniques ; Genetic Engineering ; Glucuronidase ; Orchidaceae ; genetics ; Plants, Genetically Modified ; genetics ; Polymerase Chain Reaction ; Transformation, Genetic

BACKGROUND: Animal experiments showed that recruitment maneuver (RM) and protective ventilation strategy of the lung could improve oxygenation and reduce extravascular lung water. This study was to investigate the effects of RM on respiratory mechanics and extravascular lung water index ( EVLWI) in patients with acute respiratory distress syndrome (ARDS). METHODS: Thirty patients with ARDS were randomized into a RM group and a non-RM group. In the RM group, after basic mechanical ventilation stabilized for 30 minutes, RM was performed and repeated once every 12 hours for 3 days. In the non-RM group, lung protective strategy was conducted without RM. Oxygenation index (PaO2/FiO2), peak inspiratory pressure (PIP), Plateau pressure (Pplat), static pulmonary compliance (Cst) and EVLWI of patients before treatment and at 12, 24, 48, 72 hours after the treatment were measured and compared between the groups. Hemodynamic changes were observed before and after RM. One-way ANOVA, Student's t test and Fisher's exact test were used to process the data. RESULTS: The levels of PaO2/FiO2 and Cst increased after treatment in the two groups, but they were higher in the RM group than in the non-RM group (P<0.05). The PIP and Pplat decreased after treatment in the two groups, but they were lower in the RM group than in the non-RM group (P<0.05). The EVLWI in the two groups showed downward trend after treatment (P<0.05), and the differences were signifcant at all time points (P<0.01); the EVLWI in the RM group was lower than that in the non-RM group at 12, 24, 48 and 72 hours (P<0.05 or P<0.01). Compared with pre-RM, hemodynamics changes during RM were significantly different (P<0.01); compared with pre-RM, the changes were not significantly different at 120 seconds after the end of RM (P>0.05). CONCLUSIONS: RM could reduce EVLWI, increase oxygenation and lung compliance. The effect of RM on hemodynamics was transient.

ObjectiveTo investigate the protective effect of transfected microRNA-146a (miR-146a) on mice with sepsis-induced acute lung injury (ALI) in vivo.Methods Twenty-four healthy male BALB/C mice were randomly divided into sham group, sepsis group, transfection group and transfection control group, eachn = 6. Mice in transfection group were given miR-146a agomir loaded by in vivo-jetPEITM via airway before reproduction of model, and mice in transfection control group were given negative control loaded by in vivo-jetPEITM only via airway. The septic model was reproduced by cecal ligation and puncture (CLP) 12 hours after transfection , and the mice in the sham group underwent laparotomy and closure only without ligation and puncture of the cecum. The mice of each group were sacrificed at 24 hours post-operation. The expression of miR-146a in lung tissue was determined by real time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR), and the quantity of tumor necrosis factor-α (TNF-α) in the bronchial alveolar lavage fluid (BALF) was determined with enzyme-linked immunosorbent assay (ELISA). The wet/dry ratio of lung (W/D) was determined. The pathohistological changes in the lung were observed and scored. Results The expression of miR-146a showed a significant increase in sepsis group, transfection group and transfection control group, which were (3.56±0.43), (27.64±3.46) and (3.72±0.54) folds of that in sham group, respectively (P< 0.05 orP< 0.01). The miR-146a expression in transfection group was significantly increased compared with sepsis group and transfection control group (bothP< 0.01), but no statistical difference in the expression was found between sepsis group and transfection control group (P> 0.05). Compared with the sham group, higher level of TNF-αin the BALF was found in the sepsis group, transfection group and transfection control group (ng/L: 511.65±43.47, 305.74±34.76, 492.27±42.21 vs. 50.72±7.23, allP< 0.01). The level of TNF-α in transfection group was significantly lower than that in sepsis group and transfection control group (bothP< 0.01). Compared with the sham group, the W/D ratio of lung in sepsis group, transfection group and transfection control group showed a significant increase (6.11±0.32, 5.02±0.29, 6.05±0.43 vs. 4.18±0.10, allP< 0.01). The W/D ratio of lung in transfection group was significantly lower than that of sepsis group and transfection control group (bothP< 0.01). The lung injury score of transfection group was significantly lower than that of sepsis group and transfection control group (6.12±0.75 vs. 10.53±1.52, 9.73±1.08, bothP< 0.01).Conclusions miR-146a agomir loaded by in vivo-jetPEITM instillation into airway was able to increase the expression of miR-146a in the lung tissue of septic mice. Up-regulation of miR-146a inhibit the release of the inflammatory cytokine TNF-α stimulated by sepsis, and alleviate inflammatory reaction and lung tissue injury in mice with sepsis-induced ALI.

Objective To observe the expressions of OPN, COX-2 and CyclinD1 in breast infiltrating carcinoma and evaluate their relationships with clinic pathological features. Methods Expression of the above three indexes were detected from 70 breast cancinoma patients by immunohistochemistry. The relationships among them and clinicopathological features were analyzed. Results The positive expression rates of OPN were 78.8% in cases (≤45 years old) and 73.0% in cases (> 45 years old); the positive expression rates were 79.3%(tumor diameter ≤ 3 cm) and 73.2% (tumor diameter > 3 cm); the positive expression rates were 77.8%, 73.8% and 78.9% in cases ofⅠgrade, Ⅱgrade and Ⅲ respectively, the positive rates had no statistical significances(P > 0.05). The expression rates of OPN in cases of breast infiltrating carcinoma without and with axillary node metastasis were 62.5% and 93.3%, in cases at stage Ⅰ~Ⅱ and Ⅲ ~Ⅳ were, 68.0% and 95.0% respectively, the positive rates had statistical significances(P < 0.05). The expression of OPN was negatively correlated with ER and PR while positively correlated with CerbB2, COX-2 and CyclinD1. Conclusions OPN plays an important role in the invasion and metastasis of breast carcinoma coordinated with COX-2 and CyclinD1.

AIM:To investigate the effects of glucagon-like peptide-1 (GLP-1) receptor agonist exendin-4 on white adipose tissue (WAT) and the underlying mechanisms.METHODS:Male C57BL/6J mice (8 weeks) were chal-lenged by high-fat diet for 12 weeks, and were randomly divided into saline group and exendin-4 group.The mRNA expres-sion of sirtuin 1 (SIRT1), adipose triglyceride lipase (ATGL), TNF-αand adiponectin of WAT was detected by real-time PCR.3T3-L1 adipocytes or mouse embryonic fibroblasts cells were treated with exendin-4 for 24 h.The protein levels of SIRT1, ATGL and hormone-sensitive lipase (HSL) were determined by Western blot.RESULTS:Exendin-4 significantly decreased epididymal fat weight, fasting blood glucose and serum triglyceride levels ( P<0.05) , and reduced body weight and serum TNF-αlevel.The mRNA expression of SIRT1, ATGL and adiponectin in WAT was all significantly up-regulated by exendin-4, which were contrary to the down-regulation of TNF-αmRNA expression (P<0.05).Exendin-4 promoted the protein expression of SIRT1, ATGL, and HSL in 3T3-L1 adipocytes in a dose-dependent manner.Less lipid droplets with up-regulation of lipolytic protein expression were observed when combined with SIRT1 agonist treatment, which were suppressed by SIRT1 inhibitor.Deletion of SIRT1 led to larger adipocytes with more lipid droplets, and the effect of ex-endin-4 on the lipolysis disappeared when SIRT1 was deficient.CONCLUSION:Exendin-4 promotes lipolysis in WAT of obese mice via activation of SIRT1.

Diversified teaching methods including traditional teaching,problem based learning,interactive teaching and team cooperation teaching were adopted in the process of teaching.Problem based learning can stimulate students' learning interests; interactive teaching can improve the students' hands-on ability and team cooperation teaching can cultivate students' cooperation spirits.Diversified teaching mode made different teaching methods integrated and complemented each other,forming a special teaching mode for the course of functional experiment.

Objective To evaluate the efficacy of early continuous high-volume-hemofiltration in the treatment of patients with severe acute pancreatitis (SAP).Methods Based on the method of prospective,randomized and controlled clinical trial,60 patients with SAP between January 2005 and July 2011 from the First Affiliated Hospital of Nanchang University were divided into control group and hemofiltration group.The hemofiltration group was treated with early continuous high-volume-hemofiltration and not in the control group.The changes of vital signs,clinical symptoms and laboratory indicators were compared between the two groups before and after the treatment.Results After hemofiltration,the clinical symptoms such as abdominal pain,fever,tachycardia and respiratory distress in hemofiltration group were significantly remitted compared to those in the control group (P ＜0.05).The APACHE Ⅱ score (13.3 ± 1.0 vs 14.1 ± 1.2) and the level of TBil[(20.4±11.3) μmol/L vs (28.1 ±10.9) μmol/L],creatinine[(178.7 ±71.8)μmol/L vs (215.6 ± 51.3) μmol/L],blood urea nitrogen[(10.1 ± 5.6) mmol/L vs (13.2 ± 3.8) mmol/L] and ALT[(51.3 ± 13.2) U/L vs (62.5 ±14.3) U/L] were decreased compared to those in the control group (all P values ＜0.05).The level of PaO2/FiO2(197.3 ±32.4 vs 178.3 ±31.7) was increased (P ＜ 0.05).After hemofiltration,heart rate was decreased gradually (P ＜ 0.05) in the hemofiltration group than in the control group.Mean artery pressure (mAP) increased gradually (P ＜ 0.05) in the hemofiltration group than in the control group.Conclusion Early continuous high-volume-hemofiltration has significant effects on the treatment of SAP including the improvement of clinic symptoms,the blockade of development from systemic inflammatory response syndrome (SIRS) to multiple organ dysfuction syndrome(MODS),improvement of organ function and prevention from the complications.It may become one of the important therapies for SAP.