Objective To investigate the effect of astragalus polysaccharide,a component of an aqueous extract of Astragalus Membranaceus roots,on differentiation and maturation of dendritic cells in vitro. Methods 30 BALB/c mice were randomly divided into three groups,normal control group,100,200 mg/kg APS intraperitoneal injection groups. After one week,weight the mouse spleen,account the splenetic index. Collect the mouse bone marrow cell,induced and cultured with rmGM-CSF and rmIL-4. With inverted microscope to investigate the morphous of DC cell. The phenotypes of DC were detected by flow cytometry and the expression of the GM-CSF protein in serum was tested by ELISA. Results Astragalus polysaccharide injection had obvious effects on the spleen weight of mice. The degree of CD11c and MHC-Ⅱ expression in 100 mg/kg and 200 mg/kg groups on flow cytometry were advanced significantly compared with that in normal control group,but the degree of CD80 and CD86 was not increased. And the expression of the GM-CSF protein in serum in 100 mg/kg group and 200 mg/kg group were both not increased significantly compared with nomal control group. Conclusion The intraperitoneal injection of astragalus polysaccharide could stimulate the proliferation of the pre DC in bone marrow. The angtigen presentation of DC might be enhanced,but this effects was not positive correlation with concentration of GM-CSF.

Objective To describe an alternative procedure of arthroscopic double-bundle posterior cruciate ligament (PCL)reconstruction using deep-frozen fresh allogenic achilles tendon and evaluate the follow-up outcome in early stage. Methods The ruptured PCL was reconstructed under arthroscopy using deep frozen fresh allogenic achilles tendon preparing in Y-shaped allografts, one bundle was used for anterolateral bundle, the other for posteromedial bundle. Absorbable screw was used to fix the soft tendon graft and metal screw was used to fix the bone graft. Results From March 2003 to August 2004, the procedure of arthroscopic double-bundle PCL reconstruction using allogenic achilles tendon was applied in 10 cases. The average follow-up period was 13.2 months(6～24 months, among them 6 cases were more than 12 months ). 2 cases with anterior cruciate ligament (ACL) injury were also reconstructed with hamstring tendons (semitendinosus and gracilis). 3 cases with rupture of postolateral horn of knee joint were anatomically reconstructed with hamstring tendons. Clinical results were carefully compared using the Lysholm score system before and after the operation, the average score was 50.3 preoperatively, and 91.2 postoperatively. The objective examination showed that the tibia post-migrating sign, posterior drawer test and Lachman test were positive in all the 10 cases preoperatively; whereas the posterior drawer test and Lachman test were negative postoperatively; the tibia post-migrating sign were weak positive in 3 cases with longer clinical histories(more than 1 year); the flexion range of the knee were slightly restricted (5?～20?) in the 2 cases with simultaneous ACL injury. Conclusions Allogenic achilles tendon is an ideal material for arthroscopic double-bundle PCL reconstruction. Arthroscopic double-bundle reconstructed PCL is in accordance with the anatomy and physiology of natural PCL. The injury and complications caused by the autograft can be alleviated. The allograft can be prepared in advance and the soft end of the implant can pass through the bone tunnel easily, and thus the operaton time under arthroscopy can be saved and the trauma caused by arthroscopy is abated. Arthroscopic double-bundle PCL reconstruction is sample, safe and effective.

Objective To introduce the method of two bundle anatomical reconstruction of the posterolateral corner(PLC) of the knee using auto-harmstring,and to evaluate the short-term clinical outcomes.Methods Using auto-harmstring,23 knees of 21 cases with acute or chronic posterolateral complex injuries were anatomically reconstructed from March 2003 to November 2005.There were 20 male and 1 female patients(mean age:32.3 years,ranging from 17 to 47 years)in this study with follow-up of at least 12 months(average:26.7 months,ranging from 12 to 31 months.Tow cases had isolated PLC injuries,21 knees had multiligamentous injuries,and 5 cases associated with meniscus injuries.The technique used includes:(1)an autogenous semitendinosus placed through trastibial bone tunnel from posterior to anterior and then turned to transfibulur bone tunnel to reconstruct the popliteus and popliteofibular ligament;(2)an autogenous gracilis tendon placed through transfibulur bone tunnel to reconstruct the fibulocolateral ligament(FCL).The transplanted grafts were secured to the femoral insertion of the popliteus and FCL respectively with metal or bioabsorbable interference screw.All patients were followed-up prospectively with clinical examinations and Lysholm knee scores.Results There was no varus knee instability in full extension,and one-grade varus instability with firm endpoint was found in 2 cases at 30? flexion.There was no increased external rotation in all of the 23 knees at 30? flexions(in prone position).Mean Lysholm knee scores were 89.2(range from 88 to 100).Conclusions Two bundle anatomical reconstruction of the PLC of the knee using auto-harmstring yields a stable knee with excellent function,mini-trauma,and reliable fixation of grafts,this technique is an ideal method for the treatment of PLC injuries.

Objective To study the relation between the pressure in forefoot and the second metatarsalgia in hallux valgus deformity, finally to quantitatively analyze the pressure under the second metatarsal head which is a dangerous factor leading to second metatarsalgia. Methods Three groups volunteered for this study by a self-devised foot pressure measurement system: fifty-eight normal subjects (116 feet) without pain and deformity in foot, thirty-two patients (45 feet) with hallux valgus and pain only under the second metatarsal head, fifty-six patients (102 feet) with hallux valgus without symptoms of metatarsalgia. The percentage of pressure to weight was the criteria for data analysis. First the pressures under second metatarsal head among three groups were compared with each other. Secondly we used case-control study to identify whether it was high pressure that was likely to result in metatarsalgia. Finally we compared the pressure interval under second metatarsal head among three groups to define the pressure interval which might be a dangerous factor to second metatarsalgia. Results 1)The pressure beneath the second meatatarsal head in patients with metatarsalgia was the highest in the three groups (P

Three antibodies were used for immunohistochemical study of 15 cases of adenoid cystic carcinoma(6 cases contained normal salivary gland tissue). The results indicated that myoepithelial cell of the normal salivary glands revealed positive reaction of S-100 protein; The basal cells of the straited ducts and excretary ducts revcaled strong positive reaction of keratin. In adenoid cystic carcinoma positive reaction of S-100 prctein and vimentin were found in luminal cells of cribriform pseudocyts and peripheral cells of tumor nests, and the outer layers'cells of tubular stucture and the border cells of trabucula. But the positive reaction of keratin was found in inner cells of tubular and central cells of trabecular patten. It indicated that adenoid cystic carcinoma probably stems from the basic cells of the striated ducts and excretary ducts of normal salivary glands.

AIM To evaluate the changes of serum and brain tissue endoxin in model of bilateral cerebral hemisphere ischemic reperfusion injury, and effect of anti digoxin antiserum (an antagonist of endoxin). METHODS The bilateral cerebral hemisphere ischemic model was prepared by ligating three vascular by Kameyama's manner. SD rats were randomly divided into 7 groups and each group had 8 rats. Sham group, ischemic reperfusion group, negative control group, nimodipine group, low concentration anti digoxin antiserum group, middle concentration anti digoxin antiserum group, high concentration anti digoxin antiserum group. The blood was collected at the end of reperfusion, meanwhile rats were killed, and the bilateral cerebral hemisphere were took out and used to prepare encephlon homogenate and made into samples of light microscope. RESULTS Compared with sham group, the serum CK content increased; Brain tissue SOD activity reduced and MDA content increased importantly in ischemia reperfusion group; The levels of serum and brain tissue endoxin in ischemia reperfusion group were significantly higher, while ATPase activity in brain tissue decreased; Mitochondrial Ca 2+ content in brain tissue increased significantly and Mg 2+ content decreased significantly. In brain tissue,there was some inflammatory change and local necrosis;The rank order and structure of cell wasn't clear;The morphology of pyramidal cell was abnormal. Compared with ischemic reperfusion group, Anti digoxin antiserum reduced serum CK content; It antagonized lowering of SOD activity and increase of MDA content in brain tissue; It remarkably reduced the level of brain tissue endoxin; It reduced abnormal ion content of brain tissue mitochondrion induced by cerebral ischemic reperfusion injury; The high and middle concentration anti digoxin antiserum had a significant effect on raising brain tissue ATPase activity. It reduced neuron denaturation. CONCLUSION Cerebral ischemic reperfusion can increase the level of brain tissue and serum endoxin and higher endoxin can promote brain injury. Endoxin is a major factor involved in cerebral ischemic reperfusion injury. Anti digoxin antiserum can reduce brain tissue injury and had a protective and treatment effect on cerebral ischemic reperfusion injury by antagonizing the effect of endoxin.

Objective To evaluate the efficacy and safety of dorsal root entry zone (DREZ) incision for pain induced by brachial plexus injury. Methods A cohort study was performed in 41 consecutive patients with pain induced by brachial plexus injury treated with DREZ incision from July 2005 to December 2007. Of all, 19 patients were amputated and had phantom limb sensation. Oral pain relief (0%-100%) and complications were followed up by one professional doctor. Oral pain relief rate ≥50% was considered satisfactory outcome. Results The proportion of patients with satisfactory pain relief was 90% (37/41), 81% (30/37), 77% (24/31) and 70% (16/23) at 2 weeks, 3 months, 6 months and last follow-up (over one year) after surgery. At last follow-up, 15 out of 16 patients with over 10 years of pain duration got satisfactory pain relief; while only 16 out of 25 patients with less than 10 years of pain duration got satisfactory pain relief (x2 =4.682, P =0.030). In 16 amputees who were followed up for more than three months, seven patients with alteration of phantom limb sensation got satisfactory pain relief, while only four out of nine patients without alteration of phantom limb sensation got satisfactory pain relief. Among 37 patients followed up for more than three months, severe neurological defects were found in three patients including one with mild motor disturbance in the ipsilateral lower limb and two with severe sensory disturbance in the ipsilateral lower limb. Conclusions DREZ incisions are effective and safe neurosurgical procedure for relieving pain induced by brachial plexus injury. The possible predictive factors of better outcome are the long preoperative pain duration and postoperative phantom limb sensation among amputees.

BACKGROUND:Valvular interstitial cel s are the main components of the heart valves. Myofibroblasts, as a kind of valvular interstitial cel s, can express alpha-smooth muscle actin and type I col agen fiber, and hold differentiation potential. These cel s cannot only play a support role in the valve structure, but also play a regulatory role in the process of the valve normal physiological and pathological responses.
OBJECTIVE:To obtain a reliable method of separation, primary culture and identification of myofibroblasts laying a foundation for further study on the cardiac valvular calcification.
METHODS:Aortic valve myofibroblas extracted from porcine hearts were primary cultured by trypsin and col agenase combined digestive method, common enzyme-digestion method and tissue-culture method, respectively. The myofibroblast activity and morphology were observed using microscope, and myofibroblasts were identified using light microscope and immunocytochemistrial method.
RESULTS AND CONCLUSION:Myofibroblasts had a higher activity and purity cultured by trypsin combined with col agenase II digestion method. Aortic valve myofibroblasts were positive for alpha-smooth muscle actin and negative for von Wil ebrand factor under fluorescence microscope, suggesting that myofibroblasts were successful y obtained.

Objective:To determine the effect of Hes1 on bone marrow CD34+cells in acute myeloid leukemia (AML). Meth-ods:Bone marrow mononuclear cells were isolated by using Ficoll. Then, the proportion and cell cycle of CD34+cells were analyzed by using fluorescence-activated cell sorting (FACS). CD34+cells were cultured in vitro for colony-forming cells (CFC). The expression of Hes1 in CD34+cells was evaluated by using real-time polymerase chain reaction. After upregulating the expression of Hes1 in CD34+cells, the cell cycle was analyzed through FACS, and the colony formation of CD34+Hes1+cells was analyzed by CFC. Results:The ra-tio of CD34+cells in the bone marrow was lower in the AML group than in the control group. In addition, more CD34+cells underwent quiescence in the AML group than in the control group. In vitro assay showed that the colony formation of CD34+cells was lower in the AML group than in the control group. The expression of Hes1 was higher in the CD34+cells from the AML patients than that in the CD34+ cells from normal donors. After Hes1 transduction, more CD34+ cells underwent quiescence and showed weak proliferation. Conclusion:The proportion of CD34+cells in the bone marrow was lower in AML patients than in normal donors. A large proportion of CD34+cells underwent quiescence, which was related to Hes1, in AML patients.

Objective To investigate the safety and feasibility of a new pancreaticojejunal anastomotic method [the penetrating-suture type of pancreaticojejunostomy (PPJ)] after pancreaticoduodenectomy (PD).Methods From May 2005 to November 2011,a new surgical anastomosis was carriedout between the pancreas stump and jejunal wall with penetrating-suture technique after pancreaticoduodenectomy in 46 patients.The clinical data were reviewed and analysed.Results The anastomosis of the pancreas stump and jejunal wall was completed with six to eight interrupted full-layer sutures.There was no pancreatic anastomotic leakage and hemorrhage.Conclusion The PPJ is a simple and effective approach and can be recommended.