Fluorescence resonance energy transfer(FRET)is increasingly used to study inter-and intramolecular interactions in living cells.Since being proportional to the concentration of the donor-acceptor complex.FRET value must be normalized to exclude the influence of the ratio and the concentration of donor and acceptor for comparison.Different from the intra.molecular FRET which is simplified by the fact that the COncentration of the donor is equal to that of the acceptor,the inter-molecular FRET is usually too complicated for most existing measurements to quantify exactly.We deduced the exact proportion of the donor-acceptor complex based on a unique characteristic of homodimer,a special kind of the intermolecular interaction,developed an exact quantification measurement of the FRET.We proved the novel method Can generate more reliable estimation of FRET value by comparison with other methods using a homodimer,estrogen receptor alpha(ERa),as a FRET pair.

Background Retinal vascular recanalization is key to the treatment of retinal vascular occlusive disease.Studies confirmed that urokinase by intravitreal injection inhibits the expression of occludin protein at tight junction complexes among retinal capillary endothelial cells.Objective This study was to observe the effects of urokinase via eye local injection on the outward permeability of blood-retinal barrier by detecting the concentration of intravitreal Evans blue (EB).Methods Sixty healthy Sprague-Dawley (SD) rats were randomly assigned to four groups,and the right eyes of the rats were used as experimental eyes.Urokinase of 4 μl (350 U) and the equal volume of PBS (0.01 mol/L) was intravitreally injected separately in the intravitreal urokinase group and the intravitreal PBS group,and 10 μl urokinase (1000U) and the equal volume of PBS was injected via retrobulbar tissue respectively as the retrobulbar urokinase group and the retrobulbar PBS group.Twenty-four hours after injection of drugs,0.5％ EB 4 μl was intravitreally injected.Four hours later,the rats were sacrificed and the right eyeballs were excised for the extraction and drying.EB was extracted from dried vitreous by formamide.Then,the concentration of EB in formamide was determined by a formamide extraction-ultraviolet spectrophotometry method to calculate the concentration of EB in vitreous.The use and care of experimental animals followed the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission (2011 version).Results The rat vitreous body showed the light blue color in intravitreal urokinase group and the retinal vessels were visible under the microscope,and that in the retrobulbar urokinase group presented blue color.However,in the intravitreal and retrobulbar PBS group,rat vitreous exhibited the deeper blue color and retinas were invisible.Absorbance of EB in formamide was 0.181 ±0.008,0.450±0.017,0.330±0.009 and 0.436±0.012 in the intravitreal urokinase group,intravitreal PBS group,retrobulbar urokinase group and retrobulbar PBS group,respectively.The intravitreal EB concentrations in the intravitreal urokinase group were (0.266±0.014)g/L,which was lower than (0.667±0.026) g/L,(0.496±0.015) g/L and (0.657±0.017) g/L of the intravitreal PBS group,retrobulbar urokinase group and retrobulbar PBS group,showing significant different among the four groups (F =100.406,P＜0.01),and the intravitreal urokinase group showed the lowest value in comparison with other three groups (all at P＜0.01).Conclusions Local application of urokinase around eye can augment the outward permeability of blood-retinal barrier in rats.Intravitreal assay of EB after intravitreal injection is a feasible approach to the determination of outward permeability of blood-retinal barrier.

Objective To investigate the prevention and management of portal vein thrombosis following orthotopic liver transplantation.Methods Between May 1995 to September 2005,clinical data of 137 cases subject to orthotopic liver transplantation were analyzed.Among them,there were 10 cases of portal vein thrombosis(5 cases in gradeⅠ,4 cases in gradeⅡand 1 case in gradeⅢ). All patients received an eversion thromboendovenectomy(ETEV)with occlusion of the portal flow u- sing a Forgarty balloon.Ligation of the collateral circulation,especially spontaneous or surgical sple- norenal shunt,was made as approaches to improve portal flow.Heparin or low-molecule-weight hepa- rin as a prophylactic anticoagulation therapy was maintained during and after operation if prothrombin time was less than 18 s.Follow-up Doppler ultrasonography was used daily in the early postoperative period.Results After a follow-up of 2～66 months,overall incidence of portal vein thrombosis was 2.92%(4/137).Surgical thrombectomy and revascularization was carried out in 1 case.Thromboly- sis,balloon angioplasty and stent placement via hepatic artery were performed on 2 cases.No treat- ment was given in 1 patient without hepatic dysfunction and portal hypertention.Mortality related to portal vein thrombosis was 0.Conclusions Portal vein thrombosis might be avoided by performing a complete thrombectomy as often as possible,by ligation of portosystemic shunt during surgery,and by postoperative anticoagulation.Close follow-up by Doppler ultrasonography may make a prompt di- agnosis and reduce portal vein thrombosis-derived loss of grafts.

Objective To explore the relationship between the amount of bleeding and the histopathologic grade of cerebral glioma with susceptibility weighted imaging (SWI), so that to assess the diagnostic value of SWI in grading cerebral glioma. Methods Totally 30 patients with cerebral glioma underwent SWI. The area of hemorrhage of every slice displayed in SWI was measured with software. The rate and amount of bleeding were compared between high and low grade tumors. Results There was no statistical significance in the incidence of detected bleeding on SWI (P=0.064), though it was higher in highly malignant group (80.00%) than that in low grade group (46.67%), while there was statistical significance in the amount of bleeding detected on SWI (Z=-2.275, P=0.026) between highly malignant group and low grade group. Conclusion The amount of tumor hemorrhage displayed in SWI is valuable for the preoperative grading of cerebral glioma.

Objective:To investigate the prognostic significance of plasma fibrinogen (FIB) and D-dimer in elderly patients with non-small cell lung cancer (NSCLC) before any anti-cancer treatments.Methods:97 cases of patients with lung cancer (lung cancer group) and 36 healthy subjects (control group) were enrolled;the level of plasma D-dimer and FIB were compared;the relationship between their levels and clinical pathological factors of NSCLC as well as prognosis was analyzed.Results:The levels of plasma FIB and D-dimer in lung cancer group were higher than those in healthy control group (P＜0.05).FIB in lung cancer group was related to TNM stage,and D-dimer was related to lymph node metastasis and TNM stage.Univariate analysis showed that FIB,D-dimer,tumor size,lymph node metastasis and TNM staging were associated with overall survival time (OS) and progression free survival (PFS).Multivariate analysis revealed that only D-dimer and FIB were independent prognostic factors of patients with NSCLC.Conclusion:Detection of fibrinogen and D-dimer in elderly patients with NSCLC before anti-cancer treatment would be useful for prognosis and might provide a certain value for individualized therapy.

Objective To investigate the change of levels of related cytokines,including transforming growth factor-β2 (TGF-β2),basic fibroblast growth factor (bFGF),platelet-derived growth factor (PDGF),interleukin-6 (IL-6),interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α) and behavior in streptozotocin-induced diabetic rats between the before and after fracture,at the same time,to analyse the relativity of them.Methods The rat models of diabetic were induced by intraperitoneal injection of streptozotocin,which were divided into different diabetes mellitus group (DM group) randomly.The right tibiofibula of all the rats of diabetes mellitus group and normal control group(NC group) were broken,and then treated with external fixation.The bone callus formation was evaluated by X-ray.When all the operation above was completed,the rats were domesticated in different cages.The concentration of related cytokines of the DM groups and NC group was detected at different time.Results The behavior changes of diabetes fracture rats were distinct and appeared a series symptoms such as bradykinesia,dull,polydipsia,polyphage,polyuria,emaciated obviously.The concentration of related cytokines (TGFβ2,BFGF,PDGF,IL-6,IL-8 and TNF-α) of the two groups was detected at 1,2 and 4 weeks.The difference between them was significantly(TNF-α:the concentration of NC group were (9.87 ± 1.06) ng/ml,(14.07 ± 1.43)ng/ml and(12.02 ± 1.02) ng/ml; the concentration of DM group were (11.65 ± 1.18) ng/ml,(16.29 ± 1.24)ng/ml and(13.87 ± 1.25)ng/ml ; P ＜ 0.05).X-ray films showed that the fracture healing of NC group was more superior than that of DM group.Conclusion The reduction of TGF-β2,BFGF,PDGF and the increase of IL-6,IL-8,TNF-α in early fracture are one of important reasons for delayed union and nonunion in diabetic rats.At the same time,there is a cooperative action between them.Dynamic observation of the concentration of related cytokines will provide eliable experimental data to estimate fracture healing,delayed union and nonunion.It is favourable to acquaint the regulatory mechanism of fracture healing at molecular level.

Objective To investigate the effect of hepatopoietin Cn(HPPCn) on liver stem cells.Methods In this study, WB-F344 cell line was used, and MTT and flow cytometry assay were conducted to determine cell proliferation and apoptosis.Transwell assay was used to test the migration of WB-F344 cells.A 2AAF-partial hepatectomy(PH) mouse model was used to observe the effect of HPPCn on liver stem cell proliferation in vivo.Results HPPCn enhanced WB-F344 cell proliferation and migration and activated the SphK1, Erk and Stat3 signal pathways.The analysis of the 2AAF-PH mouse model showed that oval cells in the experimental group far outnumbered those in control and the regeneration of the liver was improved post PH.Conclusion HPPCn can increase the liver stem cell proliferation and survival while promoting the regenenation of the liver by augmenting oval cell proliferation.

Objective To investigate the clinical efficacy of Jin’s Three Needles plus twelve Jing-Well points in treating post-stroke shoulder-hand syndrome. Methods Sixty patients meeting the inclusion criteria of post-stroke shoulder-hand syndrome were randomly allocated to treatment and control groups, 30 cases each. The treatment group received acupuncture with Jin’s Three Needles plus twelve Jing-Well points and the control group, conventional acupuncture. The Clinical Neurological Disfunction Scale (CNDS) score, the Fugl-Meyer Assessment (FMA) score, the Visual Analogue Scale (VAS) score and the Functiional Comprehensive Assessment (FCA) score were observed in the two groups before and after two weeks of treatment. The clinical therapeutic effects were compared between the two groups. Results There were statistically significant pre-/post-treatment differences in the CNDS, FMA, VAS and FCA scores in the two groups (P<0.05). The total efficacy rate and the cure and marked efficacy rate were 96.7%and 55.7%in the combination group, respectively, in the treatment group and 83.3%and 26.7%, respectively, in the control group;there were statistically significant differences between the two groups (P<0.05). Conclusion Acupuncture with Jin’s Three Needles plus twelve Jing-Well points is an effective way to treat post-stroke shoulder-hand syndrome.

Objective To compare the short-term effects and toxicity of one cycle of TPF regimen induction chemotherapy with that of two cycles of PF regimen induction chemotherapy in the N advanced local nasopharyngeal carcinoma (NPC). Methods A total of 47 patients initially treated with N advanced local NPC without distant metastasis from Jan 2007 to Dec 2008 were enrolled in this study. The TPF chemotherapy regimen was administered as follows: TAX 60 mg/m2, i.v. infusion on day 1st, cisplatin 80 mg/m2, i.v. infusion on day 1st and 5-Fu 800 mg/m2, bolus infusion in 96 hours on day lst-4th. The PF chemotherapy regimen was as follows: cisplatin 100 mg/m2, i.v. infusion on day 1 and 5-Fu 1000 mg/m2, bolus infusion in 96 hours on day lst-4th. Twenty-six patients received one cycle of TPF regimen and 21 patients received two cycles of PF regimen, with 21 days each cycle and a total of 3 courses in each group, then chemoradiotherapy with cisplatin 30 mg/m2 (weekly) was followed. Results The short-term efficacy was no significant difference between two groups, after one cycle TPF chemotherapy, the CR+PR rates of primary site and lymph nodes were 57.7 % and 69.2 %, respectively, while after two cycles PF chemotherapy in control group, those were 66.7 % and 71.4 %, respectively (P>0.05). In 3 months after treatment, the CR rates of primary site and lymph nodes in TPF group were 92.3 % and 88.7 %, respectively, while those in control group were 100.0 % and 90.5 %, respectively (P>0.05). The main adverse reactions were Grade Ⅲ-Ⅳ neutropenia and alopecia in TPF group. Conclusion Short-term effects of induction chemotherapy with one cycle of TPF regimen in the N advanced local NPC is satisfied, and the main toxicity is neutropenia.