A new and efficient reaction system has been set up, in which the MseⅠ primers were fluorescent labeled for auto-sequencer. PCR reagents and primers and adapters of MseⅠ and EcoRⅠ, which were synthesized, the AFLP protocol has been modified, and reaction and electrophoresis conditions were optimized, the results obtained can be comparable to that of AFLP fluorescent labeled AFLP kits with less cost.

ObjectiveTo explore the relationship between the expression of Dishevelled2 and Vangl2 and the embryonic neural tube defects (NTDs) induced by all-trans retinoic acid (RA)in Kunming mouse. Methods Fifty pregnant mice were randomly divided into control and RA-treated groups.RA-treated mice were fed with 30mg/kg RA dissolved with peanut oil on embryo 7.75 days, while the mice of control group were administrated with an equal volume of peanut oil on the same time. Then all the embryos were sampled from pregnant mice at the 4th, 18th, 42nd, 66th and 90th hour after treatment. In situ hybridization and immunohistochemical staining technique were used to detect the expression of Dishevelled2 and Vangl2 in embryonic neural tube. Results The two proteins both existed in the epithelial tissue of the mouse embryonic neural tube and displayed different expression modes at various developmental stages.Compared with the control group, the RA treated group showed a significant decrease (P≤0.05) at the 18th and 42nd hour and a significant increase (P≤0.05) at the 66th hour in Dishevelled2 protein after maternal treatment, and no significant difference was found at the 90th hour. Compared with the control group, the Vangl2 mRNA expression in the RA treated group displayed a significant decrease (P≤0.05) at the 4th and 18th hour and a significant increase (P≤0.05) at the 66th hour after RA treatment, and no difference was found at the 42nd hour. Compared with the control group, the expression of Vangl2 protein in the RA treated group decreased (P≤0.05) at the 18th and 42nd hour, and increased (P≤0.05) at the 90th hour after RA treatment, no difference was found at the 66th hour. Conclusion Excessive RA may interfere with the normal embryonic neural tube closure by regulating the expression of Dishevelled2 and Vangl2.

Objective To investigate the mammographic,galactographic,pneumographic features and their pathological basis.Methods Mammography(n=26),galactography(n=13),pneumography(n=5)and specimen radiography(n=12) in 26 cases pathologically proved,and mammographic-pathologic correlation were analyzed.Results The breast hamartomas in 26 cases were all benign,they were circumscribed and surrounded by a true capsule.The mammographic appearances could be divided into three types acording to different amount of fibroglandular and adipose tissue in tumors:prominantly fatty,prominantly fibrous(fibroglandular)and mixed fibrofatty.Conclusion A clearly containing areas of fat and fibroglandular density and a characteristic"slice of sausage"appearance can be showed on mammograms.Galactography and pneumography is of a great help to differential diagnosis of breast hamartomas.

Objective To explore the effect of postural reduction and balloon dilation on the correction of vertebral height following balloon percutaneous kyphoplasty (PKP) for osteoporotic vertebral coinpressive fracture (OVCF).Methods From June 2012 to December 2013,47 OVCF patients involving 57 fractured thoracolumbar vertebrae with intact posterior wall underwent PKP.In all patients postural reduction was performed under general anesthesia.The percentages of vertebral height loss before and after balloon dilation were compared.The curative effects were evaluated by comparing preoperative and postoperative visual analogue scale(VAS) scores and cobb angles.Results Compared with preoperation,the vertebral heights of anterior and middle columns were significantly improved after postural reduction,balloon dilation and PKP (P ＜ 0.05),but there was no significant improvement before and after balloon dilation (P ＞0.05).The postoperative average cobb angle (22.8°± 8.1°) was significantly decreased compared with the preoperative one (22.8° ± 8.1°) (P ＜ 0.05).The VAS scores at preoperation,one day postoperation and the final follow-up were respectively 8.5 ± 0.4,3.4 ± 0.2 and 3.1 ± 0.3.The postoperative ones were significantly lower than the preoperative one (F =7.518,P =0.006).Conclusions In PKP,postural reduction under general anesthesia plays an important role,but balloon dilation may exert an insignificant effect on restoration of vertebral height.

Objective To evaluate the role of β-arrestin-1 in inhibition of endotoxin-induced activation of nuclear factor kappa B (NF-κB) in human pulmonary microvascular endothelial cells (HPM-VECs) by penehyclidine hydrochloride (PHC).Methods HPMVECs were seeded in 6-well plates (2 ml/hole) or in culture flasks (4 ml/flask) at the density of 1 × 105/ml,and were randomly divided into 5 groups (n =20 each) using a random number table:empty plasmid transfection group (group C),lipopolysaccharide (LPS) + empty plasmid transfection group (group LPS),PHC + LPS + empty plasmid transfection group (group P+LPS),LPS + β-arrestin-1 gene-shRNA transfection group (group LPS+shRNA) and PHC + LPS + β-arrestin-1 gene-shRNA transfection group (group P+LPS+shRNA).HPMVECs were transfected with empty plasmid 1.5 μg or with plasmid containing 15 nmol/L β-arrestin-1gene-shRNA.At 24 h of incubation,PHC with the final concentration of 2 μg/ml was added,the cells were incubated for 1 h,LPS with the final concentration of 0.1 μg/ml was then added,and the cells were continuously incubated for another 1 h.The supernatant was collected to measure the activity of lactic dehydrogenase (LDH).The cell suspension was collected for determination of vascular cell adhesion molecule-1 (VCAM-1) expression and NF-κB activities and NF-κB inhibitor I-κB and β-arrestin-1expression.Results Compared with group C,the activities of LDH in supernatant were increased,VCAM-1 expression was up-regulated,NF-κB activity was significantly increased,and I-κB and β-arrestin-1 expression was down-regulated in LPS and LPS+shRNA groups.Compared with group LPS,the activities of LDH in supernatant were decreased,VCAM-1 expression was down-regulated,NF-κB activity was significantly decreased,and I-κB and β-arrestin-l expression was up-regulated in group P+LPS,and no significant change was found in the parameters mentioned above in group P+LPS+shRNA.Compared with group P+LPS,the activities of LDH in supernatant were increased,VCAM-1 expression was up-regulated,NF-κB activity was significantly increased,and I-κB and β-arrestin-1 expression was down-regulated in group P+LPS+shRNA.Conclusion PHC inhibits endotoxin-induced activation of NF-κB in HPMVECs completely through up-regulating β-arrestin-1 expression.

Objective To study the expression of KAI1 and CD40 in gastric cancer and their correlation with clinicopathologic features and prognosis.Methods The expression of KAI1 and CD40 of 64 gastric cancer tissues was examinated by S-P immunohistochemieal methods.Results The overall positive rate of KAI1 was 22% in gastric cancer.Expression of KAI1 was positively related to the degree of tumor differentiation,it was negatively related to invasion depth and lymph node metastasis and the tumor clinical stage.The 5-year survival rate of cases with positive expression of KAII was significantly higher than that of cases with negative expression(X~2=42.426,P=0.000).The expression rate of CD40 in gastric cancer tissue was 34%.The expression of CD40 was significantly correlated with metastasis and clinical stage of gastric cancer.The 5-year survival rate of cases with positive expression of CD40 was significantly lower than that of cases with negative expression(X~2=4.841,P=0.028).There is no relationship between KAI1 and CD40 expression in gastric cancer.Conclusion KAI1 is associated with gastric cancer differentiation,invasion and metastasis.CD40 status helps to evaluate metastasis,and predict prognosis of gastric cancer.

OBJECTIVETo discuss clinical effects of surgical treatment for desmoplastic fibroma of bone.

METHODSBetween June 2000 and June 2010, 15 cases of desmoplastic fibroma were treated by surgical operation including 4 males and 7 females with an average age of 39 years old (ranged from 18 to 64 years old). The site of tumor was proximal femur in 4 cases, distal femur in 3, distal tibia in 2, proximal humerus in 1, distal humerus in 1, scapula in 1, pelvic in 2, manubrium of sternum in 1. The simple intralesional curettage was performed in 1 case. The other 14 cases were divided into two groups, 7 cases had an aggressive curettage with inactivation and the last 7 cases had a wide resection. Recurrence condition were observed after operation. The function was valuated in two groups after the operation according to Enneking's standard.

RESULTSThe mean duration of follow-up was 56 months (ranged, 18 to 132 months). Two cases recurred, but no metastasis. The patient with simple intralesional curettage recurreed, 1 of the 7 patients with a wide resection recurred. The recurrence rate was 13.3% (2/15). There was no recurrence in the group with an aggressive curettage with inactivation. According to Enneking's standard, Enneking scoring was 21.6 +/- 3.8 in the group with a wide resection and 28.3 +/- 1.3 in another group, The results were excellent in 2 cases and good in 5 in the group with a wide resection, excellent in 7 in the other group.

CONCLUSIONThe aggressive curettage with inactivation has better functional recovery than the wide resection,and it should be chosen when the lesion is small or located in an area where reconstruction is difficult.

Adolescent ; Adult ; Bone Neoplasms ; pathology ; surgery ; Female ; Fibroma, Desmoplastic ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; epidemiology ; Young Adult

Objective To investigate the effect of apolipoprotein E gene knock-out(ApoE KO) and high-fat diet on morphology and the expression of mortalin in hippocampal CA_3 neurons of mice, and to explore the impact of these factors on memory and Alzheimer's disease.Methods Ten wild-type and 10 ApoE KO mice were fed with common chow as the control group and the KO group respectively while 10 ApoE KO mice were fed with high fat diet.Twelve weeks later, the weight and the lid of these mice were measured.The brain tissues were observed using HE staining, nissl staining, protargol staining,immunohistochemistry staining and image analysis by computer.Results In the ApoE KO group, weight,total cholesterol, triglyceride, low-density lipoprotein cholesterol were higher than those in the control group,and these changes were more significant in ApoE KO high-fat diet group.The nissl was higher in the ApoE KO group (0.301±0.031) and in ApoE KO high-fat diet group (0.261±0.020) than those in the control group (0.341±0.035, F=18.068, P<0.05).The mortalin in the ApoE KO group (0.322±0.060) and in ApoE KO high-fat diet group (0.391±0.041) were higher than the control group (0.256±0.061, F=15.230, P < 0.05).Conclusions ApoE KO and high-fat diet can reduce nissl, and improve the expression of mortalin.This protein may be involved in the pathogenesis of Alzheimer's disease.

Objective To investigate the role of p38 mitogen-activated protein kinase (p38 MAPK) in attenuation of lipopolysaccharide (LPS)-induced human umbilical vein endothelial cell injury by penehyclidine hydrochloride (PHC).Methods Human umbilical vein endothelial cells were provided by Medical Research Center,Wuhan University,cultured and seeded in 96-well plate (100 μl/hole) or 24-well plate (3 nl/hole) with density of 1 × 104/ml or in culture flasks (5 ml/flask) with density of 1 × 106/ml.The cells were randomly divided into 4 groups ( n =23 each):group control (group C) ; group LPS; group PHC (group P) and group PHC + LPS (group PL).The cells were exposed to LPS 1 μg/ml in groups L and PL or/and PHC 2 μg/ml in groups P and PL.LPS was added at 1 h after PHC in group PL.The cells were collected at 24 h exposure to LPS for determination of the expression of phosphorylated p38 MAPK (p-p38 MAPK) and p38 MAPK.The ratio between p-p38 MAPK and p38 MAPK was calculated.Cell viability,NO content and inducible nitric oxide synthase (iNOS) expression were also determined.Results LPS significantly decreased cell viability,increased NO content,iNOS expression,p-p38 MAPK and p-p38 MAPK/p38 MAPK ratio in group L as compared with group C.In group PL pretreatment with PHC significantly attenuated LPS-induced cell injury.Conclusion p38 MAPK pathway is involved in attenuation of LPS-induced endothelial cell injury by PHC.