AlM:To explore the suitable conditions in rapid model of corneal neovascularization ( CNV ) after thermal burn under different constant temperature in rabbit.
METHODS: Total 45 New Zealand white rabbits were divided randomly into five groups ( A, B, C, D, E ) . A groups:100℃ ( n = 10 ) , B groups: 200℃ ( n = 10 ) , C groups:300℃ ( n=10 ) , D groups: 400℃ ( n=10 ) , and E groups:control group ( n=5 ) . All left eyes of rabbits in A,B,C,D groups were induced corneal neovascularization by constant temperature burning device. The growth of CNV was observed by slit lamp microscope and the area of CNV were recorded on 4 th , 7 th , 14 th , 30 th days postoperatively. SPSS 19. 0 statistical package was used for data analysis, and the data was recorded by mean ± standard deviation. Comparison by analysis of variance was made by repeated measures in the area of neovascularization at each time point in groups. Statistical tests were considered significantly when P values were less than 0. 05.
RESULTS:On postoperative 4 th , 7 th , 14 th , 30 th days: no neovascularization was found after corneal thermal burn in A group, but only a few nebula left (n=2);the area of CNV were (9.16±1.45)mm2, (37.73±5.49)mm2, (62.44± 7. 54 ) mm2 , ( 40. 28 ± 7. 39 ) mm2 in B group respectively;and (11.45±1.04)mm2, (44.51±4.64)mm2, (66.13±4.13)mm2, (43.04±2.33)mm2 in C group respectively; and (13.23± 0.86)mm2,(47.26±4.59)mm2,(67.57±4.56)mm2,(45.59± 4. 44 ) mm2 in D group respectively, and part corneal carbide ( n = 4 ) was observed as well as corneal perforation ( n= 6 ) were found on 3d in D group. No neovascularization was found in normal control group. Comparison of the areas of CNV at each time point between groups was statistically different, P < 0. 05. Statistical differences were found among B, C, D groups, P<0. 05.
COCLUSlON:ln 4 to 7d, the higher the temperature is, the more the neovascularization area of CNV are. lt has no significant difference in 14 to 30d. But corneal carbide and corneal perforation are often found in 400℃ group, so its modeling failure rate is high. lt is between 200℃ and 300℃ that repeatability and uniformity of the corneal neovascularization model of rabbit are superior.

Objective:To generate rabbit polyclonal antibody against human Argonaute2 (Ago2) protein and to identify its functional characterization for determination of differential expression and cellular localization of Ago2 protein in various cell lines.Methods:DNAstar software was applied for searching the high antigenicity region of Ago2 gene sequence termed k-Ago2.Prokaryotic expressing plasmid was constructed and transformed to E.coli BL21 (DE3) to induce expression by IPTG.The fusion protein was injected into rabbits subcutaneously to produce polyclonal antibodies after purification by gel regaining.ELISA was operated to detect antibody titer.Western blot was used to identify the specificity and sensitivity of the antibodies and detect the differential expression of Ago2 protein in various cell lines.Meanwhile,immunofluorescence experiments were arranged to show cellular localization of Ago2 protein.Results:The prokaryotic expressing plasmid was constructed correctly.K-Ago2 protein was expressed and purified,and then rabbit polyclonal antibodies against Ago2 were generated after immunization with k-Ago2 protein.The titer detected by ELISA was 1∶19 000.Western blot results demonstrated the high specificity of the antibodies.Finally,we successfully observed the differential expression and cellular localization of Ago2 protein in various cell lines.Conclusion:The polyclonal antibody against Ago2 protein has been achieved successfully.It will be propitious for the intensive study of the RNAi mechanism and even profound clinical application.

Objective To study prognosis of patients with fulminant hepatitis after plasma ex- change treatment using model for end-stage liver disease(MELD)scoring system.Methods 160 pa- tients were randomly divided into plasma exchange group and control group,and MELD score was calculated according to the original formula for each patient.The efficacy of plasma exchange was as- sessed by mortality and improvement in biochemical parameters and MELD score.Results The levels of total bilirubin(TBIL),INR and MELD score of patients whose MELD scores were between 30 and 40[TBIL,(379.4?40.4)?mol/L; INR,2.5?0.2; MELD,30.8?3.8]were lower than before PE treatment[TBIL,(509.7?64.6)?mol/L;INR,3.5?0.3;MELD,37.3?3.5].The levels of TBIL and INR and MELD score of patients whose MELD scores were higher than 40 [TBIL,(595.6?61.5)?mol/L;INR,3.8?0.4;MELD,39.8?3.5]were lower than before PE treatmem [TBIL, (650.4?66.3)?mol/L;INR,4.4?0.6;MELD,45.2?4.2].The mortality of patients in PE group with MELD score from 30 and 40 was 50.0%,while it was 86.7% in control group,showing significant differ- ence between PE group and control group(P＜0.01).The mortality of patients with MELD scores higher than 40 was 91.2% in PE group and 100% in control group,showing no significant difference between these two groups(P＞0.05).Conclusions Plasma exchange treatment can decrease the serum TBIL level, INR and MELD score of patients with fulminant hepatitis and improve liver function.Compared with the control group,plasma exchange can significantly decrease the mortality of patients in PE group with MELD score from 30 to 40,but no effect on patients with MELD score higher than 40.

Background The domestic and international researches discovered that many proteins and enzymes of the extracellular matrix (ECM) participate in the sclera remodeling by affecting the collagen typeⅠand fibronectin.Objective This study was to investigate the effect of matrixmetalloproteinase-2 (TIMP-2) on expression of collagen typeⅠand fibronectin of ECM in the posterior sclera by injecting liposomes containing tissue inhibitor of TIMP-2 gene into suprachoroidal space of the form-deprivation myopia in guinea pig.Methods Form-deprivation myopia was induced by translucent goggles in 36 clean guinea pig for 2 weeks.Then the animals were randomly assigned to TIMP-2 group,empty plasmid group,saline group and 12 for each group.Liposomes of 5μl containing TIMP-2 gene,empty plasmid and saline were suprachoroidally injected in the right eye respectively,and the left eyes without any treatment were used as self-control group.Other 12 matched guinea pigs only covered the right eyes through out the experimental duration as model control group.The guinea pigs were sacrificed and the posterior sclera tissue of the eyeballs were collected at 2,7 and 14 days after injection of drug.The expressions of collagen typeⅠmRNA and fibronectin mRNA were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR).This study followed the Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results The expression level of collagen type Ⅰ mRNA in the posterior sclera of guinea pig was lower but that of fibronectin mRNA was higher in TIMP-2 group than self-control group,showing significant differences between them (P＜0.05).The expression level of collagen type Ⅰ mRNA in the posterior scleral tissue began to increase from the 2nd day after drug injection and was obviously elevated at the 7th day and then gradually decreased at the 14th day.However,the expression level of fibronectin mRNA in the posterior scleral tissue showed the opposite pattern.The expression levels of collagen typeⅠmRNA and fibronectin mRNA at the 7th after drug injection were significantly lower than that at the 2nd day or 14th day (P＜0.01).Conclusion Suprachoroidal injection of TIMP-2 in form-deprivation myopia could up-regulate the expression of collagen typeⅠmRNA and down-regulate the expression of fibronectin mRNA in the posterior scleral tissue.It may slow down the sclera remodeling of form-deprivation myopia in guinea pig in the early stage.

Objective The role of long non-coding RNA Linc00467 in human lung adenocarcinoma is not yet clear.This study was to investigate the expression of long non-coding RNA Linc00467 in human lung adenocarcinoma, its clinical significance, and the effects of Linc00467 on the functions of the tumor and endothelial cells in vitro.Methods Lung adenocarcinoma tissue and normal tissue surrounding the malignance were obtained from 60 patients with pathologically proved stage I-Ⅲa lung adenocarcinoma.Human umbilical vein endothelial cells (HUVECs) were transfected with the over-expressed plasmid pccl-Linc00467 (HUVEC experimental group) or the empty vector pccl (HUVEC control group), A549 cells with Linc00467-siRNA (A549 experimental group) or negative siRNA (A549 control group), and H1299 cells, too, with Linc00467-siRNA (H1299 experimental group) or negative siRNA (H1299 control group).The expression level of Linc00467 in the lung adenocarcinoma tissue was detected by qRT-PCR with an analysis of its correlation with the clinicopathological characteristics of the patients;the influence of Linc00467 on the proliferation of the A549, H1299 and HUVEC cells was assayed with CCK-8;and the role of Linc00467 in the angiogenesis of the HUVECs was assessed by fibrin bead sprouting assay.Results The expression of Linc00467 in the lung adenocarcinoma tissue was 2.72±1.31 times as high as that in the normal lung tissue (P<0.01), and those in the A549 and H1299 cells were 3.45±0.25 and 3.22±0.33 times as high as those in the human bronchial epithelial (HBE) cells (P<0.01).The expression level of Linc00467 was significantly correlated with the tumor size and vascular invasion (P<0.05).After transfection of Linc00467-siRNA, the expressions of Linc00467 in the A549 and H1299 experimental groups were down-regulated by 72% and 68% as compared with those in the A549 and H1299 control groups (P<0.01).The number of living cells was remarkably decreased in the A549 experimental group in comparison with the A549 control at 48 h (1.29±0.07 vs 1.51±0.09), 72 h (1.53±0.15 vs 2.13±0.11), and 96 h after culturing (1.98±0.18 vs 3.02±0.12), and so was it in the H1299 experimental versus the H1299 control group, but markedly increased in the HUVEC experimental versus the HUVEC control group (P<0.05).At 5 days, HUVEC experimental group, as compared with the HUVEC control, showed a significantly increased number of newly formed vascular branches (7.36 vs 4.25/superbead, P<0.01) and relative length of the blood vessels (3.12 vs 1, P<0.01).Conclusion Linc00467 promotes tumor cell proliferation and angiogenesis and is highly expressed in the lung adenocarcinoma tissue, which is correlated with the tumor size and vascular invasion and suggests that Linc00467 could be a potential biomarker and therapeutic target.

Objective To investigate the impact of fatty liver and its related factors on the effi- cacy of antiviral therapy in patients with chronic hepatitis C.Methods ninety-eight patients with chronic hepatitis C were treated with 180?g peginterferon?-2a plus ribavirin.Hepatitis C virus (HCV) genotypes were measured by enzyme-linked immunosorbent assay (ELISA) and fatty liver were detected by ultra sonography.The body mass index (BMI),waist to hip ratio (WHR) and ho- meostasis model assessment of insulin resistance (HOMA-IR) were calculated.The serum HCV RNA level was determined by polymerase chain reaction (PCR) and serum insulin level was detected by chemiluminescence analysis.The impact of fatty liver and its related factors on the efficacy of antiviral therapy were analyzed by multivariate Logistic regression analysis.Results Of 98 patients with chro- nic hepatitis C,35 (35.7%) were genotype 1,41 (41.8%) were genotype 2,13 (13.3%) were gen- otype 3,9 (9.1%) were undetermined genotype.The incidence of fatty liver in HCV infection of genotype 1,2,3 and undetermined genotype was 11.4%,9.8%,38.5% and 11.1%,respectively (X~2=7.83,P

Background Researches have showed that the extraocular muscle injection of insulin-like growth factor(IGF) results in the thickness of extraocular muscle and enhancement of contractility,and lack of IGF probably is associated with pathogenesis of strabismus.If extraocular muscle injection of insulin-like growth factor binding protein 4(IGFBP-4),an inhibitor of IGF,can further ensure the effect of IGF is still under-investigation.Objective The aim of this study was to evaluate the effects of IGFBP-4 on the developing extraocular muscle.Methods IGFBP-4 of 2 μl(1 g/L)was injected into superior rectus muscles via supraorbital margin in the left eyes of 24 postnatal l-day-old chickens once every other day for five times,and the equivalent amount of normal saline solution was used in the same way in the fellow eyes as controls.The animals were sacrificed by overdose of anesthesia 2 days after the final injection and 12 animals were randomly selected for the maximal contractility test of superior rectus muscle.The other 12 pieces of superior rectus muscle of the chickens were obtained for the hematoxylin & eosin staining to evaluate the morphology of superior rectus muscle and calculate the myofiber diameter using imaging system.Results The maximal contractility was (1.602 ± 0.080) mN and(1.815±0.O10)mN in the IGFBP-4 injection group and normal saline solution group respectivily,showing a significant difference between the two groups(t =13.65,P＜0.01).The maximal contraction force in cross-sectional area was (95.500 ± 7.420) mN/cm2 in the IGFBP-4 injection group and it was significantly lower than that of the normal saline solution group(101.510±5.220)mN/cm2(t =28.81,P＜0.01).Histological examination showed that thin myofibers were obvious more in the IGFBP-4 injection group than those with saline injection.The mean diameter was (8.7 ± 0.5) μm in the IGFBP-4 injected group,and that of the normal saline group was(9.1 ±0.4) μm,there was no significant difference between them (t =0.75,P =0.46).However,significant differences were found in the number of different diameters of myofibers between the two groups(all at P＜0.05).Conclusions IGF is one of the key nutrition factors in the development of extraocular musele.The extraocular muscle injection of IGFBP-4 may cause the dysfunction and morphologic abnormality of extraocular muscle.

ObjectiveTo investigate the effect of hyperbaric oxygen (HBO) on proliferation and differentiation of endogenous neural stem cells after acute spinal cord half cut-off in rats. MethodsThe differences of proliferation and differentiation of endogenous neural stem cells between injured group and intervention group were compared. ResultsThere were remarkable differences between injured group and intervention group. ConclusionHBO can promote the proliferation and differentiation of the neural stem cells in rats after spine cord injury.

Fibrosis ; Hematoma, Subdural ; Humans ; Osteogenesis ; Subdural Space ; Tomography, X-Ray Computed

[ Objective] To explore the variation features of contrast enhanced ultrasound ( CEUS) quantitative parameters and its correlation with tumor microvessel density ( MVD ) , after the treatment of living liver cancer using high intensity focused ultrasound ( HIFU ) . [ Methods ] A group of 30 New Zealand white rabbits were made into VX2 liver cancer models, that were randomly divided into control group ( n =15, not HIFU treatment) and treatment group ( n =15, HIFU treatment).CEUS was performed, then obtaining ascending slope(AS),arrival time(AT),time-to-peak(TTP),peak intensity ( PI) ,area under curve ( AUC ) .And correlation analysis was performed with MVD obtained by immune histochemical method. [ Results ] Compared with those in the control group, AS, PI, AUC were significantly decreased in treatment group (P <0.05) and so was MVD (P <0.05).AT,TTP were significantly delayed than those in the control group (P<0.05).The AS, PI, AUC of CEUS quantitative parameters were positively correlated with MVD (P<0.05).The AT, TTP respectively showed a negative correlation with MVD ( P<0.05) . [ Conclusion] CEUS quantitative parameters of liver cancer tissue have high correlation with MVD.It can reflect the microvessel structure change of the liver cancer tissue and help to judge the curative effect and prognosis of HIFU for liver cancer.