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

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.

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

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.

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.

Background To optimize the culture method of human retinal microvascular endothelial cells is very important for the study of retinal angiogenesis disease.Human retinal microvascular endothelial cells have been successfully cultured in previous studies,but further improvement of the culture method to harvest higher yields and purity cells is still needed.Objective This study was to design a modified method to isolate and purify human retinal microvascular endothelial cells much easily and quickly,and to compare the expression of specific markers of vascular endothelial cells,factor Ⅷ and CD31/CD34 in the cells.Methods The use of human donor eyeballs was approved by the Ethic Commission of Zhongshan Ophthalmic Center of Sun Yat-sen University.The retina tissue from healthy donor was isolated and digested by the two-step digestion method with 2％ trypsin and 0.133％ collagenase Ⅳ.Human retinal microvascular endothelial cells were collected and plated in 60 mm dishes coated by 0.1％ fibronectin and cultured in endothelial cell-specialized medium supplemented with 10％ fetal bovine serum,0.3 mg/L β-endothelial cell growth factor (ECGF) and 100 ng/L sodium heparin.During the culturing,the growth situation of the cells was monitored by morphological observation,and immunohistochemical staining was performed to probe vascular endothelial cell-specific membrane protein CD31,CD34 and factor Ⅷ for identification of the cell purity.Results Human retinal microvascular endothelial cells were isolated successfully from the retina by the twostep digestion method.The primary cultured cells adhered to well 72 hours later and achieved confluence with the typical cobblestone appearance 9 to 10 days after cultured.The cells exhibited the blue nuclei and reddish cytoplasm by regular haematoxylin and eosin stain and showed a strong positive response for CD31,CD34 and factor Ⅷ by immunohistochemistry.The positive dye of CD31 and CD34 was lower than Ⅷ factor in both endothelial cells.Conclusions Modified culture method of human retinal microvascular endothelial cells can improve cell culture result and purify target cells.

Objective To investigate the effect of insulin on D_5 dopamine receptor expression and function in renal proximal tubule (RPT).Methods Immortalized RPT cells and D_5 receptor transfected HEK293 (HEK-D_5) cells were used in the study to investigate the effect of insulin on D_5 receptor expression and function,and those effects were compared in RPT cells from Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR). The function of D_5 receptor was determined by measurement of the Na~+-K~+-ATPase activity in HEK-D_5 cells. Results Insulin increased D_5 receptor protein expression in a concentration and time-dependent manner in WKY RPT cells,but not in SHR.The basal level of D_5 receptor expression was higher in WKY cells than that in SHR cells. Stimulation with fenoldopam(D_1-like dopamine receptor agonist) inhibited the Na~+-K~+-ATPase activity;pretreat- ment with insulin increased the inhibitory effect of fenoldopam on Na~+-K~+-ATPase activity in HEK-D_5 cells. Conclusion The abnormal regulation of insulin on D_5 receptor expression and function might be involved in the path- ogenesis of essential hypertension.

Objective:To study the effects of pioglitazone on atherosclerosis on ApoE-/-mice,and to investigate the roles of adiponectin and its receptors.Methods:ApoE-/-mice were fed with high-fat chow for the induction of atherosclerosis and were divided into three subgroups:placebo(n=10),low-dose[10 mg/(kg?d),n=10] pioglitazone therapy,and high-dose[20 mg/(kg?d),n=10] pioglitazone therapy.C57BL/6J wild type mice(n=9) were used as control.Aortic atherosclerosis and intima-media thickness(intima-media thickness,IMT) of abdominal aorta were monitored,and plasma adiponectin was also measured.Expression levels of the adiponectin receptor 1(AdipoR1)and adiponectin receptor 2(AdipoR2) in vessels were analyzed(RT-PCR).Results:(1) Aortic atherosclerotic lesions were observed in ApoE-/-mice but not in wild type mice.Interestingly,these lesions were significantly prevented by high-dose pioglitazone therapy.Compared with wild type mice,ApoE-/-mice had increased IMT of abdominal aorta [(0.290?0.063 vs 0.178?0.012) cm,P

Objective To study the treatment effect and the mechanism of high volume hemofiltration(HVHF)on acute lung injury(ALl)induced by endotoxin in dogs.Methods Sixteen healthy hybrid male dogs were injected LPS(650?g/kg)via central vein within 30 minutes.After model establishment,all animals were divided into two groups randomly( n=8).One group received the treatment of HVHF,while another group received routine treatment.PH,PaO_2,PaCO_2 in arterial blood were recorded at O h after LPS model establishment and 4h after HVHF.Contents of TNF-?,IL-6,and IL- 10 in plasma were measured by radioirnmunity,mRNA expression of TNF-?,IL-6,and IL-10 in lung tissue homogenate were measured by RT-PCR and NF-?B activity by flow cytometer.Results After injection of LPS,PaO_2 and PaO_2/FiO_2 began to decrease,and PaO_2/FiO_2 value was