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 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.

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.

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.

Adenoma, Pleomorphic ; metabolism ; pathology ; surgery ; Adipose Tissue ; pathology ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Male ; Membrane Proteins ; metabolism ; Metaplasia ; pathology ; Middle Aged ; Parotid Gland ; metabolism ; pathology ; surgery ; Parotid Neoplasms ; metabolism ; pathology ; surgery ; S100 Proteins ; metabolism

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 observe the effect of combined therapy of external point application and herbal concentrate-granules on patients with asthma (hot wheezing in TCM).Methods 60 patients suffering from onset period of bronchial asthma were randomized to a control group and a treatment group.The control group was given budesonide inhaler and theophylline sustained release tablets.The treatment group was given combined therapy of external point application and herbal concentrate-granules.The course of treatment was 10 days.To observe the value ofFEV1％,ACT scores and TCM syndrome scores etc.Results After treatment,the value of FEV 1％ [treatment group:(82.83 ± 11.35) ％,control group:(83.85 ± 16.72) ％] and ACT [treatment group:(19.86±2.32),control group:(19.66±2.54)] in both groups were markedly increased with statistical significance (P＜0.01); and the difference between the two groups were also significant; TCM syndrome scoring [treatment group:(4.27±3.65),control group:(5.05±4.14)] was notably decreased in both groups compared with the values before the treatment with statistical significance (P＜0.01); but the difference between the two groups was not significant (P＞0.05).Conclusion The combined therapy of external point application and herbal concentrate-granules can treat bronchial asthma in the acute clinical course and improve lung function.

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.

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

OBJECTIVETo compare two kinds of method for treating lumbar tuberculosis with psoas abscess, to provide reference for clinical reasonable select of therapy treatment.

METHODSFrom January 2010 to January 2013,42 patients with lumbar tuberculosis combined with psoas abscess with obvious surgical indications were enrolled, including 24 males and 18 females with an average age of (38.5 ± 10.2) years old ranging from 21 to 63 years old. All patients were followed up for 18 to 24 months with an average of 20.9 months. Twenty-two patients underwent posterior vertebral body lesions cleared, bone graft fusion and internal fixation and percutaneous puncture catheter drainage for treatment of psoas major abscess as group A, and twenty patients underwent one-stage extraperitoneal approach to remove abscess, posterior vertebral body lesions cleared, bone graft fusion and internal fixation as group B. The operative time, loss of blood, length of hospital stay, clinical cure rate and other clinical results for the two groups were analyzed and compared.

RESULTSThe loss of blood was (452.3 ± 137.6) ml in group A and (603.5 ± 99.6) ml in group B, there was significant statistical difference (P < 0.05). The time of operation was (193.6 ± 91.2) min in group A and (230.5 ± 56.6) min in group B, there was significant statistical difference (P < 0.05). The time of operation and the loss of blood in group A were obviously less than which in group B. In group A 20 cases were cured and 2 cases relapsed, 19 cases were cured and 1 case relapsed in group B, there was no significant statistical differences between two groups regarding cure rate with chi-square test (χ² = 0.000, P = 1.000). All patients in two groups obtained good clinical curative effect. There were no significant statistical difference between two groups regarding for length of hospital stay with t-test (P > 0.05).

CONCLUSIONLumbar spinal tuberculosis with psoas abscess is not absolute indications for anterior open operation. Compared with the combined anterior and posterior surgical procedure, the percutaneous puncture catheter drainage combined with posterior debridement, interbody fusion and internal fixation can achieve the same clinical effect but less trauma for the patients.

Adult ; Case-Control Studies ; Debridement ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Psoas Abscess ; etiology ; surgery ; Spinal Fusion ; Tuberculosis, Spinal ; complications ; surgery ; Young Adult