Aim A method of coculture of brain capillary endothelial cells (BCECs) and astrocytes of rats was used to evaluate nanoparticle' s blood-brain barrier (BBB) transcytosis and toxicity at the endothelial tight junction. Methods A lipophilic fluorescent probe, 6-coumarin, was incorporated in poly(ethyleneglycol)-poly (lactide) nanoparticle using double emulsion/solvent evaporation method. BCECs and astrocytes were firstly isolated from brain of newborn rats and characterized by their morphology and immunocytochemistry staining, separately. Subsequently, a coculture model with BCECs on the top of micro-porous membrane of cell culture insert and astrocytes on the bottom side was established. The permeability of 14C-labeled sucrose and nanoparticle were determined, separately. Results The meanweight-based diameter of 6-coumarin loaded nanoparticles was ( 102.4 ± 6.8) nm, with zeta potential of ( - 16.81 ± 1.05) mV. BCECs were positive for factor Ⅷ staining and glial fibrillary acidic protein was tight junction between BCECs in the coculture model could be visualized by both scanning electron microscopy and transmission electron microscopy. The unchanged paracellular transport of sucrose proved vivo situation for examination of the permeability of nanoparticle and toxicity evaluation.

Objective To investigate the effects of methylation of p73 gene on pathogenesis of acute lymphoblastic leukemia.Methods The restriction endonuclease enzyme digestion combined with PCR was used to detect p73 gene and methylation in 42 patients with acute lymphoblastic leukemia.Results Methylation of p73 gene was found in 28.6%(12/42).Patients without methylation of p73 gene had a higher complete remission rate of first chemotherapy and a longer survival time than those with methylation of p73 gene.Conclusion Methylation of p73 gene probably plays a role in the pathogenesis of acute lymphoblastic leukemia and may have clinical significance in predicting prognosis of ALL.

Objective To investigate the expression and significance of hypoxia-inducible factor-1α (HIF-1α) in the retinal neovascularization by metabolic acidosis in newborn rats. Methods One hundred and twenty newborn SD rats were randomly divided into acidosis (experiment) and normoxia (control) groups. A total of 60 newborn rats in experiment group underwent tubal feeding day for 6 days and followed by a period of recovery. The rats in the two groups were sacrificed at the 3rd, 5th, 8th, 10th, 13th and 20th day after birth, respectively. The morphologic changes of retinal vessels were estimated by observing the vascular pattern in adenosine diphosphatase stained retina flat mounts. The newborn vessels were quantified by HE staining. Immunohistochemical method was used to detect HIF-1α expression. Results In experiment group, numerous neovascularization and un-perfused area at the periphery of vessels occurred on the 10th day. The result of HE staining showed that in experiment group of 10-day old,the number of neovascular nuclei extending into the vireo was 28.78±7.53, and that of the control group was 1.22±1.48 (t=11.169,P<0.01). The results of immunohistochemistry revealed that the expression of HIF-1α protein were stronger in the experiment group than in the control group on the 8th, 10th and 13th day, and there were significant differences between the two groups (108.87±15.21, 183.68±26.58 and 129.42±9.85 vs 74.98±4.50, 76.38± 3.38 and 74.78±1.86, t=4.625, 9.023 and 9.672,P<0.05). Conclusions HIF-lα might play an important role in retinal neovascularization.

Stroke is one of the severe perioperative complications.The incidence of perioperative stroke has great difference because of the surgical site,surgical complexity,and different basic diseases of patients.The induced anesthesia and the intraoperative management also have an important influence on the onset of stroke.The mortality of perioperative stroke is as high as 26%.Neturologists must guide the assessment of the potential risks for perioperative stroke,and carefully balance the risk-benefit ratio when making each decision.This article reviews the advances in research on the risk factors for perioperative stroke,prevention,and treatment.

Objective To explore the dynamic expression of ROCK-Ⅰ,p-MBS Thr-697,α-SMA protein and their mRNA in the hepatic fibrogenesis and the changes of actin cytoskeleton.Methods ROCK-Ⅰ and p-MBS Thr-697 protein in liver were determined by Western blot and their mRNA was examined by reverse transcription-polymerase chain reaction (RT-PCR),while the distribution of ROCK-Ⅰ and α-SMA in liver was assessed immunohistochemis-tically.The change of actin cytoskeleton was shown by fluorescence.Results With the development of hepatic fi-brosis,the positive areas in model groups at week 1 to 4 of ROCK-Ⅰ and α-SMA of the rat livers were larger than that in control group respectively(P <0.05).ROCK-Ⅰ,p-MBS Thr-697 ,α-SMA protein and mRNA were increased than that in control group respectively.ROCK-Ⅰ mRNA expression correlated with α-SMA (r =0.718,P <0.05).With the development of liver fibrosis,the images of fluorescence were inhanced.Conclusion With the develop-ment of liver fibrosis,both protein and mRNA of ROCK-Ⅰ increased.

90%inasinglestep,andtheproteinyieldwas2.5mg/L.ConclusionPurifiedChsp60kDantigenisobtained,andtheantigenmightbeappliedtodetectantibodyinpatientsinfectedwithChlamydialtrachomatis,andwillcontributetostudytheroleofChsp60inimmunopathogenesis.

Objective By observing the changes in NGF and its receptor P75 in liver of CCl4-induced toxic rats and to evaluate the role of NGF in the mechanism of hepatic fibrosis.Methods The expressions of NGF mRNA and its receptor P75 were detecteded by both hemi-quantitative RT-PCR and Western blot analysis.Results The expressions of NGF mRNA and its receptor P75 in liver of CCl4-induced toxic rats at 24th hour group were higher than that in control group(P

The professional care by multi-disciplinary team and priority of prevention should be carried out in the treatment of diabetic foot disease to reduce diabetic amputation.This article describes the professional experience in the treatment of four complicated cases with diabetic foot disease and emphasizes the importance of the co-operation among different specialists,including diabetologists and wound,vascular,orthopedic surgeons,etc.as well as of varied therapies applied in staged management of the diabetic foot care,by treating these patients with diabetic foot disease as early as possible.

Objective To identify the gene mutations of platelet membrane glycoprotein Ⅱ b,Ⅲa(GPⅡb/Ⅲa)in three Chinese pedigrees with Glanzmann thrombastIlenia.Methods All exons and exonintron boundaries of GP Ⅱ b/Ⅲ a gene were amplified by PCR analysis followed by DNA sequencing.DNA sequencing was used to exclude gene polymorphisms.Results The probands in the three pedigrees had a normal platelet count,coagulation profiles,scattered platelets on the blood film,a prolonged cutaneous bleeding time,and impaired or minimal ex vivo platelet aggregation in response to ADP,thrombin,collagen,adrenaline and arachidonic acid,but normal platelet aggregation in response to ristoeetin.Both FACS and Western blotting demonstrated trace content of αⅡb in the platelets from proband 1 and proband 3,who were classified as type Ⅰ GT,and a small amount of αⅡb in the platelets from proband 2,who was classified as type Ⅱ GT.Compound heterozygous mutations,T2255G(Leu721Arg)and C2671T(Gln860Stop)were identified in proband 1.The proband 2 had homozygous A2334C(Gln747Pro)missense mutation.Nonsense mutations C1750T (Arg584Stop)and 69-79 deletion mutation were identified in proband 3. Conclusions Compound heterozygous mutations T2255G and C2671T of αⅡb gene lead to type Ⅰ Glanzmann thrombasthenia for proband 1. Homozygous mutation A2334C of αⅡb gene leads to type Ⅱ Glanzmann thrombasthenia for proband 2. Compound heterozygous mutations C1750T and 69-79del αⅡb gene lead to type Ⅰ Glanzmann thrombasthenia for proband 3. T2255G,C1671T and 69-79del aye novel mutations for αⅡb gene.

Objective To investigate the expression of DcR3 mRNA in cells of acute leukemia(AL) and its clinical significance.Methods The DcR3 mRNA of bone marrow mononuclear cells(BMNC) in the patients with AL was detected by RTPCR.There were 28 AML cases,and 18 ALL cases.27 cases of non-malignant hematopathy were used as control group.The expression level of DcR3 mRNA in BMNC was detected with RT-PCR.Results The positive rate of DcR3 mRNA was 67.4% and gene expression level was 0.56?0.09 in AL group,while they were 40.7% and 0.39?0.19 in control group,there were significant differences(P0.05).The positive expression level of DcR3 mRNA increased when the white blood cell count was ≥30?109?L-1.Conclusion DcR3 gene might play a role by suppressing apoptosis in leukemogenesis.