A simple assay for detection of phospholipase C (PLC) activity was developed based on a fluorescence liposome probe using the Liss Rhod PE-loaded phospholipid liposomes.The liposome probe was prepared by the coassembly of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and fluorescent lipid (Liss Rhod PE).The probe showed very low background fluorescence due to fluorescence self-quenching effect of Liss Rhod PE.As the PLC enzyme selectively digested lipid, the Rhod fluorescence was recovered from its quenched state, leading to the sensitive detection of PLC.This assay provided a limit of detection (at a signal-to-noise ratio of 3) of 2 U/L for PLC.In the presence of PLC inhibitor, the fluorescent response of the sensor for PLC decreased, indicating that the assay could also be used for screening PLC inhibitors.

Objective To investigate the inhibitory effects of IBI302 on experimental choroidal neovascularization (CNV).Methods Affinity of IBI302 to vascular endothelial growth factor (VEGF) family cytokines (including VEGF-A165,VEGF-A121 and placental growth factor PlGF) and complements (C3b,C4b) was determined by enzyme-linked immunosorbent assay (ELISA).The antagonist effect of IBI302 on VEGF was measured by proliferation,migration and tube formation tests of human umbilical vein endothelial cells (HUVEC).The anti-complement activity of IBI302 was measured by hemolysis test mediated by complement classical pathway and alternative pathway.Rhesus laser-induced CNV model was divided into 5 groups including model control group,bevacizumab group,IBI302 0.25 mg group,IBI302 0.50 mg group and IBI302 1.25 mg group.Fluorescein angiography and optical coherence tomography were performed on these monkeys at 14 and 28 days after drug delivery to observe the fluorescein leakage area and retinal thickness.The aqueous VEGF concentration was measured at 29 days after drug delivery.Results IBI302 showed good affinity to VEGF-A165,VEGF-A121 and PlGF,as well as C3b and C4b.IBI302 significantly inhibited the proliferation,migration and tube formation of HUVEC induced by VEGF-A165.IBI302 inhibited the hemolysis induced by complements obviously.At 14 and 28 days after drug delivery,the area of fluorescein leakage and retinal thickness in IBI302 0.25 mg group,IBI302 0.50 mg group,IBI302 1.25 mg group were reduced.The differences of the area of fluorescein leakage and retinal thickness in three IBI302 groups were not significant (P＞0.05).At 29 days after drug delivery,the VEGF concentration in the aqueous of rhesus monkey in bevacizumab group [(38.644 ± 6.521) pg/ml] was decreased than that in model control group [(94.203± 17.360) pg/ml],the difference was significant (P＜ 0.05).The VEGF concentration in the aqueous of rhesus monkey in three IBI302 groups were less than 31.300 pg/ml.Conclusion IBI302 inhibited experimental CNV through blocking the activity of VEGF and complement.

Objective To analyze the serum keratin 18 (K18) level of patients with hepatitis B virus (HBV)-related acute-on-chronic liver failure (HBV-ACLF) and its correlation with prognosis.Methods From December 2012 to October 2014,120 patients who visited Fuzhou Infectious Diseases Hospital and diagnosed with HBV-ACLF were enrolled,and 20 chronic hepatitis B (CHB) patients and 20 healthy controls were enrolled with matched gender and age.Enzyme linked-immunosorbent assay was used for the detection of K18 (M30,M65) and the differences of M30,M65,M30/M65 and other laboratory results were analyzed among the first enrolled 40 patients with HBV-ACLF,20 CHB patients and 20 healthy controls.All HBV-ACLF patients were followed up for 3 months and divided into two groups (death group and survival group).And M30 and M65 in both groups were detected and their clinical data were collected.Continuous variables between groups were compared using t test and one-way analysis of varicmce.Categorical variables were compared using X2 test.Receiver operating characteristic (ROC) curve was used for prognostic analysis.Results The serum M30 in HBV-ACLF group,CHB group and healthy controls group were (2.99 ±0.29),(3.12±0.26) and (2.16 ±0.12) lg U/L,respectively (F=95.36,P＜ 0.01).The serum M65 in the three groups were (3.41 ± 0.29),(3.38+0.29) and (2.01±0.11) lg U/L,respectively (F=217.60,P＜0.01).And M30/M65 the three groups were 0.39 ± 0.11,0.55 ± 0.09 and 1.45 ± 0.34,respectively (F=202.63,P＜0.01).The white blood cell,total bilirubin,alanine aminotransferase and aspartate aminotransferase in HBV-ACLF patients were the highest,and platelet,albumin,serum sodium and prothrombin time activity were the lowest.After following up for 3 months of 120 patients with HBV-ACLF,40 of them died with fatality rate of 33.3％.The area under ROC curve (AUC) of the M30/M65 was 0.871,and that of score of model for endstage liver disease (MELD) was 0.668 (Z=3.011,P＜0.01).The M30/M65 ratio was significantly negatively correlated with the MELD score (r=-0.389,P＜0.01).Conclusions K18 (M65 and M30)are strongly associated with liver disease severity.The M30/M65 ratio may be a potential diagnostic and prognostic marker for the patients with HBV-ACLF.

AIM:To explore the preventing effect of citalopram on neuro-cell apoptosis caused by long-term stress in CA1 and CA3 region of hippocampus.METHODS:Forty male Sprague Dawley(SD)rats were randomly divided into five groups including blank group,control group(the control group was filled the stomade by 0.9% saline)and three experimental groups(intragastric administration of citalopram hydrobromide at doses of 8 mg?kg-1?d-1,4 mg?kg-1?d-1,1 mg?kg-1?d-1,respectively).Rat stress model was made by compulsory swimming everyday for 4 weeks.Cell apoptosis in CA1 and CA3 region of hippocampus was observed by HE staining method.Apoptotic cell numbers and integral optical density in CA1 and CA3 region were tested and analyzed by terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling(TUNEL)method and Norton Internet Security BR(NIS BR)software.t-test was applied to compare apoptosis cell numbers and integral optical density.RESULTS:Control rats showed more static time and less struggling times.Conversely,static time was shorter and rats spent more time after exhaustive exercise,and more struggling times in the experimental group.Rats in control group showed more positive cells in CA1 and CA3 regions and higher integral optical density in CA3 region than those in blank group.Rats in experimental groups showed fewer positive cells in CA1 and CA3 regions.Rats in experimental group 1 and group 3 showed higher integral optical density in CA1 and CA3 regions than that in control group.CONCLUSION:Long-term stress might cause neuro-cell apoptosis in CA1 and CA3 region of hippocampus.Citalopram might have prophylactic effect on apoptosis caused by long-term stress in CA1 and CA3 region,and the prophylactic effect might not be influenced by citalopram.Our study suggests that the treatment mechanism of citalopram in neural and mental illness by long-term stress may involve in a major role by antagonizing neuronal apoptosis in both the CA1 and CA3.

Objective To describe and examine the clinical characteristics and spectrum of tufted angiomas (TA) in infants so as to explore the therapeutic approaches to Kasabach-Merritt phenomenon (KMP).Methods The clinical and follow-up data of 24 patients with TA were retrospectively analyzed between Jan.2009 and Mar.2013.The median age of the patients was 7.5 months(ranged from 18 days to 2 years),including l0 male and 14 female.Surgical excision and observation were chosen according to the lesion and conditions of the patients.The mean follow-up period was 3.6 years (1.2 to 5.4 years).The changes in the patients' s condition were established by evaluating platelet counts,and the size of lesion.Results Common clinical features included dusky red or violaceous infiltrating cutaneous lesion,thrombocytopenia,pain or decreased function and hyperhidrosis or hypertrichosis.The following 3 clinical patterns of TA progress were described:spontaneous complete or partial regression (n =2,8.3％),TA without complications and persistence over the years(n =9,37.5％),and TA complicated by KMP(n =13,54.2％).The average interval of development of KMP for delayed cases was 45.2 days(ranged 0 d to 4 months).Each of the 13 patients who developed KMP subsequent to initial presentation was symptomatic at the time KMP was detected(enlarged lesion,n =8;increased lesion firmness with change in cutaneous stain,n =3 ;and respiratory distress,n =2).All of 13 patients were cured by surgery.Complete surgical resections were performed on 10 cases.The thrombocyte count was back to normal within 1-3 days post operation,and hemoglobin and blood coagulation function gradually returned to normal within 1 to 2 weeks.Other 3 cases received major resection surgery.The number of platelets in the patients were unstable,but significantly higher than that of preoperational stage.The platelet count remained above 60 × 109/L.The residue lesions in 2 cases disappeared gradually after the operation and medication were given within 3-6 months.And the other case died of multiple organ failure post-operation.Conclusions Surgical intervention can be applied to TA that severely makes damage to children's appearance or looks or may be complicated with KMP or functional abnormality.A closely monitored policy seems appropriate for the early small tumor without severe complications.And it is necessary to monitor the number of the platelet regularly and find the KMP by as early as possible.TA associated with KMP is vitol to infants.And surgical therapy after clear diagnosis should be done as early as possible.The surgical therapy is a reliable management with higher curative rate,short disease period and minimal side-effect.

AIM:This study continuously monitors the hemodynamic changes in conscious and anesthetic rats during rapid ascent to high altitude to investigate whether there is difference between the 2 conditions and discuss the rela-ted underlying mechanism.METHODS: Sprague-Dawley rats were randomly divided into conscious group, anesthetic group, anesthetic-5000-control ( A-5000-control) group, anesthetic-5000-aminoguanidine ( A-5000-AG) group, conscious-5000-control ( C-5000-control ) group and conscious-5000-aminoguanidine ( C-5000-AG ) group.The rats in anesthetic group and conscious group were kept in a hypobaric chamber, in which the simulated altitude was increased from 2 260 m to 5 000 m at 2 m/s, and the rats in other 4 groups were at 5 000 m.The system arterial pressure ( Psa) , central venous pressure ( CVP) , heart rate ( HR) and breathing rate ( BR) were directly and continuously displayed and digitally recorded by a high-performance data acquisition (PowerLab 16/35, AD Instruments) at 200 Hz.RESULTS: The HR and BR in the conscious rats were higher and MAP was lower than those in the anesthetic rats obviously.A significant decrease in mean arterial pressure ( MAP) in conscious and anesthetic groups was observed following the increase in the altitude levels, and the net decrease in MAP in conscious group was significantly greater.Additionally, HR in the conscious rats was sig-nificantly lower at 5 000 m than that of the initial level.The rats in C-5000-AG group and A-5000-AG group showed a sig-nificant increase in the arterial pressure after the intravenous injection of AG, a selective inhibitor of inducible nitric oxide synthase ( iNOS) , and no marked change of HR and BR was found.CONCLUSION: Blood pressure and HR decrease during rapid ascent to high altitude, while the change of BR is not obvious.The mechanisms of self-safety would be trig-gered in the early stage of hypoxia, which activates iNOS and then leads to a larger number of nitric oxide.Plentiful NO di-astolizes the vessels to improve the ventilation-perfusion mismatch and lower the blood pressure.When the altitude arise to 5 000 m, even more earlier, a decompensatory stage may occur in the body, leading to decreased HR and blood pressure further more than those in the anesthetic rats.Due to the effects of pentobarbital sodium, the depression of blood pressure requires a lag period and the net decrease in MAP is less than that in the conscious rats.Therefore, hemodynamic changes during rapid ascent to high altitude in conscious rats are more comprehensive and authentic.

Objective To investigate the status of depression with anxiety symptoms, and analyze the influencing factors of anxiety symptoms from demographic data and social psychological factors. Methods Hamilton depression rat?ing scale (HAMD), Hamilton anxiety rating scale (HAMA), Eysenck personality questionnaire (EPQ), life event scale (LES), trait coping style questionnaire (TCSQ) and social support scale (SSS) were used to evaluate 729 patients with de?pression. According to HAMA scores, patients were divided into non anxiety symptoms group (HAMA<7) and anxiety symptoms group (HAMA>14). Social psychological factors were compared between two groups, and the influencing fac?tors of anxiety symptoms were analyzed. Results The incidence of anxiety symptoms in depression was 58.85% (429/729), and 119 cases (16.32%) were certainly without anxiety symptoms. Compared with the group without anxiety symp?toms, the anxiety symptoms group had higher scores on neuroticism, psychoticism, negative life events and negative cop?ing style (P<0.001), but lower scores on introversion and extroversion (P=0.010). Degree of depression (OR=9.255, 95%CI:4.726~18.127), neuroticism (OR=1.595, 95%CI:1.197~2.125), negative life events (OR=1.009, 95%CI:1.001~1.017) and negative coping style (OR=1.046, 95%CI:1.013~1.080) were the risk factors of anxiety symptoms (P<0.05). Conclu?sion The incidence of anxiety symptoms in patients with depression is high. Patients with higher degree of depression and typical neurotic personality experiencing more negative life events and those with tendency to adopt negative coping style are more susceptible to anxiety symptoms.

Objective To evaluate the effect of penehyclidine hydrochloride on the damage to the non-ventilated lung in the pediatric patients undergoing one-lung ventilation (OLV).Methods One hundred and twenty pediatric patients of both sexes,aged 2-6 yr,with body mass index of 17-24 kg/m2,of American Society of Anesthesiologists physical status Ⅰ or lⅡ and New York Heart Association class Ⅰ or Ⅱ,undergoing elective lobectomy performed via video-assisted thoracoscope,were randomly divided into 2 groups (n=60 each) using a random number table:control group (group C) and penehyclidine hydrochloride group (group P).At 10 rmin before anesthesia induction,penehyclidine hydrochloride 0.05 mg/kg was injected intravenously in group P,and the equal volume of normal saline was given in group C.At 5 min after drug intervention (T0),immediately after onset of OLV (T1),at 60 min of OLV (T2),immediately after the end of OLV (T3),at the end of surgery (T4),and at 24 h after surgery (T5),venous blood samples were collected for determination of serum tumor necrosis factor-alpha (TNF-o),interleukin-6 (IL-6) and IL-8 concentrations by enzyme-linked immunosorbent assay.The specimens of normal lung tissues around the lung lobe to be resected were obtained at T1 and T3 for determination of the injured alveolus count (with a light microscope) and cell apoptosis (using TUNEL) and for examination of the ultrastructure of epithelial cells (with a transmission electron microscope).The injured alveolus rate (IAR) and apoptosis index (AI) were calculated.Results Compared to the value at T0,the IAR and AI were significantly increased at T3,the serum TNF-α,IL-6 and IL-8 concentrations were significantly increased at T2-5 (P＜0.05),and the pathological changes were obvious in the two groups.Compared to group C,the IAR and AI were significantly decreased at T3,the serum TNF-α,IL-6 and IL-8 concentrations were significantly decreased at T2-5 (P＜0.05),and the pathological changes were significantly reduced in group P.Conclusion Penehyclidine hydrochloride can attenuate the damage to the non-ventilated lung in the pediatric patients undergoing OLV,and the mechanism is probably related to inhibition of systemic inflammatory responses and cell apoptosis in lung tissues.

Objective To evaluate the effects of dexmedetomidine pretreatment on extracellular sig?nal?regulated kinase ( ERK) pathway during acute lung injury in a rat model of liver transplantation. Meth?ods Sixty male Sprague?Dawley rats, weighing 235-250 g, were divided into 4 groups ( n=15 each) u?sing a random number table: sham operation group (group S), liver transplantation group (group LT), low?dose dexmedetomidine pretreatment group ( group LD ) and high?dose dexmedetomidine pretreatment group ( group HD) . In LT, LD and HD groups, the model of orthotopic liver transplantation was estab?lished, and the operation time was about 4 h. Dexmedetomidine 2?5 and 5?0μg·kg-1 ·h-1 were intrave?nously infused for 1 h starting from 1 h prior to clipping the hepatic artery and portal vein in LD and HD groups, respectively. The rats were sacrificed after the end of operation, and the lungs were removed for determination of wet to dry weight ratio ( W∕D ratio) , cell apoptosis and expression of ERK mRNA, ERK, phosphorylated ERK ( p?ERK) , Bcl?2 and Bax in lung tissues and for examination of the pathological chan?ges ( with light microscope) and ultrastructure of lung tissues ( with transmission electron microscope) . The
injured alveolus rate ( IAR) , apoptosis index ( AI) and ratio of Bcl?2 to Bax expression ( Bcl?2∕Bax ratio) were calculated. Results Compared to group S, the W∕D ratio, IAR, AI, expression of ERK?1 mRNA, ERK?2 mRNA, p?ERK, Bcl?2 and Bax and Bcl?2∕Bax ratio were significantly increased in LT, LD and HD groups ( P<0?05) . Compared to group LT, the W∕D ratio, IAR and AI were significantly decreased, the expression of ERK?1 mRNA, ERK?2 mRNA, p?ERK and Bcl?2 and Bcl?2∕Bax ratio were significantly increased, and the expression of Bax was significantly down?regulated in LD and HD groups (P<0?05). Compared to group LD, the W∕D ratio, IAR and AI were significantly decreased, the expression of ERK?1 mRNA, ERK?2 mRNA, p?ERK and Bcl?2 and Bcl?2∕Bax ratio were significantly increased, and the ex?pression of Bax was significantly down?regulated in group HD ( P<0?05) . The pathological changes of lung tissues were significantly attenuated in LD and HD groups as compared with group LT, and in group HD as compared with group LD. Conclusion The mechanism by which dexmedetomidine pretreatment mitigates cell apoptosis during acute lung injury is related to activation of ERK pathway in a rat model of liver trans?plantation.

Objective To investigate the intervention effect of Feikang Granules (FG) on airway remodeling and D-xylose excretory rate in pulmonosplenic asthenia rats with chronic obstructive pulmonary disease (COPD). Methods SPF SD rats were equally randomized into four groups:normal control group,COPD model control group,high-dose FG group,and low-dose FG group. The pulmonosplenic asthenia rats models with COPD were established by the way of smoking with cigarettes,repeated infusion of lipopolysaccharide in the airway and feeding with decoction of Folium Sennae.The general health state of rats was observed,and pathological changes of lung tissue were examined by the optical microscope. The wall thickness of small airway in rats was measured and the excretory rate of D-xylose was detected by bromoaniline method. Results Compared with the model control group,weight was increased,the pathological changes of pulmonary alveoli andbronchiole were relieved,small airway wall thickness became thin,and D-xylose excretory rate increased in high-dose and low-dose FG groups and the normal control group,the difference being significant (P