Objectives:To develop a preparative method of continuous acid-urea-polyacrylamide gel electrophoresis (CAU-PAGE) to purify the defensin components from human and rabbit neutrophils (HNP-1～3 and RNP-1～5).Methods:Neutrophils were isolated from human peripheral blood or rabbit artificial aseptic ascites.The components of defensin were extracted by 5% acetic acid directly followed by elution of CAU-PAGE,and purified HNP-1～3 or RNP-1～5 were harvested.Polyacrylamide gel filtration was used as the method for comparison.Results:For each elution circa 1.50 mg of purified HNP or 2.50 mg of RNP were obtained from 5×108 of neutrophils,and their natural biological activity was preserved.Conclusions：CAU-PAGE is a relatively simple,rapid,economical method for purifying cationic peptides as the components of defensin.

Objective To investigate the association between the lumbar vertebral fracture damage degree with the fracture classification,injury score,kyphosis deformity and nerve function injury.Methods According to the damage degree of posterior lig ament complex(PLC),the patients were divided into the PLC intact group and PLC injury group.Its relationship with PLC injury was researched by evaluating the fracture classification,injury score and nerve function injury situation in the two groups.Results The LCS score and TLICS score in the PLC injury group were (7.1±0.8) points and (8.2±0.6) points,which were higher than (5.7±0.5) points and (4.6±0.7) points in the PLC intact group.The Denis score in the PLC injury group was more serious.The Cobb angle in the PLC injury group was 29°,and which in the PLC intact group was 19°.The proportion of nerve function insufficiency in the PLC injury group was 89％,while which in the PLC intact group was only 60 ％.Conclusion The thoracolumbar vertebral fracture is closely related with PLC.PLC damage degree is positively correlated with the fracture classification,injury score,kyphosis deformity and nerve function injury degree.

Objective To determine the anti-SARS coronavirus (SARS-CoV) activity of defensins from human and rabbit, and explore the effects of defensin during the onset and progression of SARS. Methods Defensins were purified from neutrophil of human and rabbit respectively, namely HNP (human neutrophil protein) 1-3 and RNP (rabbit neutrophil protein) 1-5, by acid-urea-polyacrylamide gel electrophoresis (CAU-PAGE). The effect of defensin on Vero-E6 cell infected by SARS CoV BJ01 strain as well as the replication of virus were observed by micro-culture technique. Rusults The HNP 1-3 and RNP 1-5 separated by CAU-PAGE presented a single band with molecular weight of approximately 4000 dalton on 15% SDS-PAGE. HNP 1-3 was identified and validated by quantitative ELISA. The activity of anti-escherichia coli (ATCC25922 strain) of HNP 1-3 and RNP 1-5 was tested by MIC determination method, and the IC50 values obtained were 25～50?g/ml and 5～10?g/ml, respectively. In the antiviral experiment, HNP 1-3 and RNP 1-5 in the concentration from 25 to 100?g/ml were not able to prohibit cytopathic effect (CPE) of SARS-CoV and the replication of virus in Vero-E6. Conclution The natural defensins (HNP 1-3 and RNP 1-5) purified from neutrophils do not play any prohibitory or eliminative effects on SARS-CoV, indicating that that human and rabbit do not have natural immunity against SARS-CoV.

AIM:To investigate the expression and the significance of VEGF-C/D in rat cornea after alkali burning as well as the role of lymphangiogenesis in the high-risk corneal transplantation rejection.
●METHODS:The model of alkali burn corneal was made. Different times corneas were taken to electron microscope for vascularization, and examined the expression of VEGF-C/D and VEGFR-3 in l, 3, 5, 7, 14, 28d. The other rat cornea after alkali burn were divided into four parts to penetrate keratoplasty, containing only blood vessels in the cornea ( group A ) , angiogenesis and lymphangiogenesis ( group B ) , lymphangiogenesis degenerating period ( group C ) , angiogenesis degenerating period ( group D ) . ln addition, there are also normal groups ( group N ) to compare the Rl values and survival time of corneal graft.
●RESULTS: Electron microscopy showed that, when the first 7d rat cornea appeared neovascularization after alkali burn, but not lymphangiogenesis. The occurrence of new blood vessels and lymphatic in 2wk. There were no obvious lymphangiogenesis in 5wk and the angiogenesis gradually subside in 8 wk. The expression of VEGF-C/D and VEGFR-3 in the corneas of rats were up-regulated in the third days after the injury, and reached its peaks at 5d. The average survival time of group N, A, B, C, D were (14.25±0.62)d, (9.35±1.02)d, (5.06±1.13)d, (8.71±0.83) d, (9. 44±1. 05)d after transplant cornea. Compared to the rest of the group, group B plant average survival time significantly shortened (P<0. 05), while compared with group B, the survival time of A, C, D groups were significantly longer (P<0. 05).
● CONCLUSION: VEGF - C/D and VEGFR - 3 are expressed significantly after corneal alkali burn. New lymphatic vessels can accelerate high - risk corneal transplantation immune rejection.

Objective To develop a PCR-SSP method for detection of HLA-DRB1 alleles in the patients who were hypersensitive to Platanus Acerifolia pollen allergen,and to probe into the association between the atopic subjects to Platanus Acerifolia pollen allergen and HLA-DRB1 alleles.Methods DNA in whole blood was extracted by phenol-chloroform method.Eight pairs of specific primers for alleles were synthesized,and HLA-DRB1*0401,*0402,*0403,*0404,*0405,*0406,*0407,*0408 alleles in 20 atopic patients and 36 healthy individuals of Jiangsu Province with Han nationality were detected by PCR-SSP(polymerase chain reaction-sequence specific primer).Results By optimizing the experimental conditions PCR-SSP methods for detection of the 8 alleles were established and the distributing data of above-mentioned HLA DRB1 were obtained.The frequency of HLA DRB1*0405 and *0406 in the patients group was higher than that of in healthy controls group,while the frequency of HLA DRB1*0402 in the patients group was lower than that in controls.No significant deference for the other 5 alleles was found between the 2 groups.Conclusion HLA-DRB1*0406和*0405 seems to be the likely suspected candidate alleles responsible for susceptibility to Platanus Acerifolia pollen allergen in the atopic patients,while DRB1*0402 might be contribute to the related resistance to the allergen.

The authors tried to combine the theoretical learning with experimental and clinical teaching by adding teaching materials of public opinion,head-simulator,typical cases,clinical practice,examination of clinical operation skill,and make up with deficiency to some extend in current pattern of stomatology teaching. The results have showed that the pattern of experimental teaching not only raises the students'activity,consolidates their basic knowledge,develops their clinical thinking ability,but also increases their ability of clinical operational skill before clinical training.

Adolescent ; Adult ; Aged ; China ; epidemiology ; Dust ; Female ; Humans ; Lung Diseases ; epidemiology ; prevention & control ; Male ; Middle Aged ; Occupational Diseases ; epidemiology ; prevention & control ; Pneumoconiosis ; prevention & control ; Young Adult

Objective To investigate the effects of electro-acupuncture on gastric emptying and Fos expression in the neuron of vagal-solitary complex (VSC) of rat bulbus after lipopolysaccharide (LPS) intraperitoneal injection (i.p.). Methods 40 male SD rats were randomly assigned to 4 groups: control group, LPS i.p. group, LPS i.p. plus electro-acupuncture at Tsusanli point group and LPS i.p. plus electro-acupuncture at non-meridian-non-acupoint group, 10 rats for each group. Immunohistochemical techniques were used to detect the Fos expression in VSC. The animal's gastric emptying was measured by phenol red method. Results The rats with gastric emptying decreased greatly to (20.7±4.5)% 2.5 hours after LPS injection, and Fos-positive neurons were significantly found in VSC (83.2±6.6) compared with control group. However, in the group of LPS i.p. plus electro-acupuncture at Tsusanli point, the gastric emptying was up-regulated to (44.1±6.2)%, and the expression of Fos -positive neurons were down-regulated to (37.9±3.8) compared with LPS i.p. group. No significant difference was found between the group of LPS i.p. plus electro-acupuncture at non-meridian-non-acupoint and the group of LPS i.p. Conclusion LPS i.p. can retard the gastric motility in rats, electro-acupuncture at Tsusanli point may well regulate the function in LPS model rats. This function may be connected with its protective effects on Fos immunoreactive neurons activity in VSC of rat bulbus.

Objective To discuss the anatomical and radiographic parameters of medullary screw fixation of anterior acetabular column so as to provide reference for clinical application. Methods Thirty cadaveric pelvic specimens (including 18 males and 12 females) were involved in the study and fixated re-spectively with 30 retrograde medullary cannular lag screws through pubic tubercle to acetabular posterosu-perior on the left side and with 30 anterograde medullary cannular lag screws through acetabular posterosu-perior to pubic tubercle on the right side. Then, two-dimensional multiplanar CT reconstruction was done to measure the angle and length of the anterograde and retrograde medullary screw trajectory, the relationship of screws with acetabular bone and penis and determine optimal point and angle of screw insertion. Re-suits The entrance of the retrograde medullary screw was located at pubic tubercle, with vertical dimen-sion of ( 17.15±1.82) mm to pubic symphysis and that of (20.51±2.19) mm to superior margin of pu-bis. For the anterograde medullary screw, the distance from the entrance of the screw to greater sciatic notch was (33.25±2.35) mm, with safe insertion angle of (32.1±2.7)°of cephalon tilting at the sagittal plane and (46.5±3.6)°of lateral tilting at the coronal plane. The entrance angle of the anterugrade and retrograde medullary screw trajectory was similar, with only opposite direction and minor difference between the left and the right sides (P>0.05). The length of medullary screw trajectory was (119.5±2.2) mm, with insignificant difference between both sides ( P >0.05). The maximum diameter of the screw was 7.2 mm. Conclusions Medullary screw fixation of anterior acetabular column is clinically feasible but needs rather high accuracy and can be used as an alternative to plate fixation.

BACKGROUND:Though the effects of conservative or traditional open reduction and internal fixation in the treatment of thoracolumbar fractures are reliable and satisfactory for most cases, two methods also have shortages. Minimal-invasive percutaneous pedicle screw system provides a new available method. OBJECTIVE:To investigate the clinical effect of Sextant percutaneous pedicle screw system in the treatment of thoracolumbar fractures. METHODS:A total of 55 patients, who had undergone percutaneous pedicle screw fixation using Sextant system (25 patients) or traditional open internal fixation (30 patients) for single-level vertebral body compression fractures in Department of Orthopedics of Renmin Hospital of Wuhan University from February 2011 to January 2013, were enrol ed in this study. RESULTS AND CONCLUSION:Except two patients in traditional open internal fixation group were lost after discharge, al other patients were fol owed up for 8-14 months. Operative time, intraoperative blood loss, postoperative drainage amount and hospital day were better in percutaneous pedicle screw fixation group than in the traditional open internal fixation group (P<0.05). Compared with preoperation, the sagittal Cobb angle, visual analogue scale and Oswestry disability index after operation were significantly lower (P<0.05), while anterior vertebral body height ratio was significantly higher in each group (P<0.05). No significant difference was found in terms of correction loss and Oswestry disability index between two groups at 8 months after operation (P>0.05). The results show that percutaneous pedicle screw fixation using Sextant system has a satisfactory outcome in the treatment of thoracolumbar fractures. However, obeying indication strictly is very important for clinical application.