Objective To establish effective method for large-scale purification of islet cells from pig pan-cress. Methods Pig pancreas tissue was digested with collagenase P followed by purification in a HCA-Fi-coil dis continuous gradient using Cobe2991 cell separator. After isolation, the islet cell yield and purity were evaluated with light microscope with DTZ staining, and the islet function assessed by insulin release as-say in vitro. Results The number of the islets coll ected from each pancreas averaged (275 000±20 895)islet equivalents (IEQ) before purification, and (230 350±26 679) IEQ after the purification with discon-tinuous gradient centrifugation. From each gram of the pancreatic tissue, (2710±229) IEQ were obtained with an average purity of (50.2±1.95) %. The purified islets responded well to high-concentration (16.7 mmol/L) glucose stimulation with a 4. 74-fold increase of insulin secretion over the basal level (3.3 mmol/L, P <0.001). Conclusion The established method can be applicable for large-scale purifi-cation of fully functional islet cells from pig pancreas.

Objective:To explore the protective effect of ellagic acid on the acute liver injury induced by carbon tetrachloride (CCl4) of the mice,and to explore its possible mechanism.Methods:A total of 50 mice were randomly divided into normal control group,model group,low,medium and high doses of ellagic acid groups (n=10).The mice in normal control group and model group were treated with 1% sodium carboxymethyl cellulose solvent by intragastic administration,and the mice in ellagic acid groups were treated with 160,320,and 480 mg·kg-1 ellagic acid by intragastic administration,respectively.After 14 d,the mice in model group and different doses of ellagic acid groups were intraperitoneally injected with 10 mL·kg-1 0.1% CCl4,while the mice in normal control group were intraperitoneally injected with 10 mL·kg-1plant oil.16 h later,all the mice were sacrificed and the body weights and the liver indexes of the mice were detected;the serum levels of alanine transaminase(ALT),aspartate transaminase(AST) were detected;the activities of superoxide dismutase(SOD) and levels of GSH-Px,malonalde hyde(MDA) and catalase(CAT) in liver tissue of the mice were detected.Results:There were no significant differences of the body weights of the mice between each group before and after treatment(P>0.05).Compared with normal control group,the liver indexes and the levels of serum ALT and AST of the mice in model group and different doses of ellagic acid groups were significantly increased (P<0.05).Compared with model group,the liver indexes of the mice in different doses of ellagic acid groups were decreased(P<0.05);the serum levels of ALT and AST of the mice in high dose of ellagic acid group were significantly decreased (P<0.05),while the CAT level in liver homogenate was significantly increased (P<0.05);the levels of GSH-Px in liver homogenate of the mice in medium and high doses of ellagic acid groups were significantly increased (P<0.05);the activities of SOD in liver homogenate of the mice in different doses of ellagic acid groups were significantly increased (P<0.05),and the MDA levels were significantly decreased (P<0.05).Conclusion:The ellagic acid has the protective effect on acute chemical liver injury in the mice induced by CCl4 and the most effective dose is 480 mg·kg-1;its mechanism may be related to the anti-oxidation.

OBJECTIVETo explore the therapeutic effects of internal fixation with Herbert screws for the treatment of Pipkin type I and type II femoral head fractures.

METHODSFrom January 2008 to December 2012, 23 patients with Pipkin type I and type II femoral head fractures were treated with open reduction and internal fixation by Herbert screws through Kocher-Langenbeck approach. Twenty-three patients (aged 18 to 52 years with a mean of 35.5 years, including 18 males and 5 females patients, 8 left hips and 15 right hips) with femoral head fractures and posterior hip dislocation. The fracture was classified according to Pipkin classification based on the radiographic findings, 5 patients had type I and 18 had type II fractures. The duration time from admission to the operation ranged from 6 to 72 h (averaged 32 h). The clinical and radiographic outcomes of the patients were measured using Thompson-Epstein scoring scale. The Harris hip score(HHS) was used to evaluate and compare hip functions at the latest follow-up between affected and healthy sides.

RESULTSAll the patients were followed up, and the duration ranged from 20 to 48 months (averaged 30 months). According to Thompson-Epstein system, 12 patients got an excellent result, 6 good, 4 fair and 1 poor. The average HHS at the finial follow-up was 87.80 ± 8.46 (ranged from 66 to 95), which is similar to that in the healthy side 90.10 ± 6.35 (ranged from 72 to 98) (t = 1.044, P = 0.302). The complications such as deep infection, and deep vein thrombosis were not found. At the 3rd year during follow-up,4 patients had avascular necrosis of femoral head. At the 1st year of follow-up, 1 patient had hip pain after walking,screws loosening and shift after trauma,and serious complications of traumatic arthritis. All the 5 patients were treated with total hip arthroplasties.

CONCLUSIONThe treatment of internal fixation with Herbert screws through the Kocher-Langenbeck approach is effective for Pipkin type I and type II femoral head fracture. The method is reliable and valuable for recommendation. However, such fracture may have avascular necrosis of femoral head and complication of traumatic arthritis, which should be observed carefully in clinic with preparation of the prevention and treatment measures.

Adolescent ; Adult ; Bone Screws ; Female ; Femoral Neck Fractures ; surgery ; Fracture Fixation, Internal ; adverse effects ; methods ; Humans ; Male ; Middle Aged

Objective To determine the effects of endostatin on the expression of vascular endothelial growth factor receptor?2 ( VEGFR?2 ) in non?small cell lung cancer cells ( human A549 lung adenocarcinoma cells and human Calu?1 lung carcinoma cells) , and to investigate the possible mechanisms underlying its radiosensitizing effect. Methods The CCK8 method was used to determine the inhibitory effect of endostatin on cell proliferation and calculate the drug concentration that caused a 20% reduction in cell proliferation within 24 h ( IC20 ) . RT?PCR and Western blot assays were used to assess the mRNA and protein expression of VEGFR?2, proteins within its related signaling pathways, and HIF?1α, respectively. The radiosensitivity of cells in each group was determined by colony formation assay;cell apoptosis and cell cycle distribution were determined by flow cytometry. Comparison of mean values between multiple samples was made by one?way analysis of variance, and comparison of mean values between two samples was made by t test. Results Endostatin significantly inhibited the proliferation of Calu?1 cells ( F=50?36,P<0?01) with an IC20 of 296?5 μg/ml;the mRNA and protein expression of VEGFR?2 and HIF?1α was also significantly inhibited in endostatin?treated Calu?1 cells ( F=25?43,10?44, all P<0?05) . Moreover, the phosphorylation of Akt, ERK 1/2, and p38 was significantly reduced in endostatin?treated Calu?1 cells ( F=2?89,0?24, 1?09, all P<0?05) . The radiosensitivity enhancement ratios for Calu?1 cells and A549 cells were 1?38 and 1?09, respectively. Endostatin significantly induced apoptosis ( F=44?15, P<0?01) and G2/M blockage ( F= 104?24, P< 0?01 ) in Calu?1 cells. Conclusions Endostatin induces apoptosis and enhances radiosensitivity in Calu?1 cells with high expression of VEGFR?2, but it has a limited impact on A549 cells with low expression of VEGFR?2.

Objective To assess the value of 11 C-methionine (MET) PET/CT for the diagnosis of residual or recurrent glioma in comparison to 18 F-FDG PET/CT. Methods Forty-six patients suspected of residual or recurrent glioma underwent both 11 C-MET and 18 F-FDG PET/CT within 5-day interval. The glioma was considered as positive on PET/CT images based on ( 1 ) visual judgment of higher tracer uptake compared with the normal gray matter; (2) semiquantitative analysis of tumor to contralateral normal gray matter (T/G) and contralateral normal white matter (T/W) ratios. The diagnosis was confirmed by stereotsctic biopsy or radiological findings of MRI or CT and clinical follow-up ( ＞6 months). The Chi-square test and paired t test were used for statistical analysis. Results Residual or recurrent glioma was confirmed in 36 patients. The sensitivity, specificity, accuracy of 11C-MET and 18F-FDG PET/CT were 94.4% (34/36) vs 47.2% (17/36), 90.0% (9/10) vs 100% (10/10), 93.5% (43/46) va 58.7% (27/46) (x2 =19. 429, 1. 053, 15. 294, P ＜0.001, ＞0.05, ＜0.001 ), respectively. The T/G and T/W ratios of residual/recurrent giloma by 11 C-MET were significantly higher than those by 18 F-FDG (T/G ratio: 1.68 ± 0.23 vs 1.13 ±0.51, t = 5. 877, P ＜ 0. 001; T/W ratio: 2.52 ± 0.28 vs 1.42 ± 0. 57, t = 10. 470, P ＜0. 001 ). Conclusion 11 C-MET PET/CT is more sensitive and accurate than 18 F-FDG PET/CT for the detection of residual or recurrent glioma.

Objective To investigate the effects of vascular endothelial growth factor receptor?2 ( VEGFR?2) on proliferation, migration, invasion, and apoptosis after radiotherapy in lung cancer cell line Calu?1, and to explore the probable mechanisms. Methods Small interference RNA ( siRNA )?mediated silencing of VEGFR?2 gene was performed on Calu?1 cells, and the mRNA and protein expression of VEGFR?2 was determined by quantitative real?time PCR and Western blot, respectively. The cells were divided into control group, vascular endothelial growth factor ( VEGF ) group, VEGFR?2 specific siRNA (siKDR) group, and siKDR+VEGF group. The changes in proliferation, migration, and invasion were evaluated by the CCK8 assay, cell scratch wound?healing assay, and transwell migration assay, respectively. The protein expression of VEGFR?2 and proteins in the related downstream signaling pathway was measured by Western blot. Apoptosis in each group was determined after radiotherapy. Results After RNA interference?mediated silencing of VEGFR?2, the mRNA and protein expression of VEGFR?2 was significantly reduced ( P=0. 001,P=0. 000);the proliferation, migration, and invasion of Calu?1 cells were also significantly reduced ( P=0. 000,P=0. 000,P=0. 000);the phosphorylation levels of AKT, ERK 1/2, and p38 were significantly reduced in Calu?1 cells ( P=0. 336,P=0. 986,P=0. 553);the apoptosis in Calu?1 cells was significantly elevated ( P=0. 0012);the protein expression of HIF?1α was significantly inhibited ( P= 0. 016 ) . Conclusions The VEGFR?2 gene silencing significantly inhibits several physiological functions of Calu?1 cells and elevates the apoptosis rate after radiotherapy.

OBJECTIVETo develop an HPLC method for the determination of eleutheroside B in As1 (the extract fraction from Acanthopanax senticosus, which has a good effect of antifatigue).

METHODThe antifatigue effect of Asl was evaluated by mice burden swimming testing. One compound was isolated by silica gel, Sephadex LH-20 column chromatography and preparative high performance liquid chromatography. The structure was identified by physicochemical properties and spectral evidences. Eleutheroside B in Asl was determined by HPLC. Chromatographic conditions included Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) and the mobile phase consisting of a mixture of acetonitrile-water-ethanoic acid (10: 90: 0.01). The UV detection wavelength was set at 344 nm.

RESULTAs1 showed an excellent antifatigue activity; One compound was isolated from As1 and its structure was identified as Eleutheroside B; The calibration cure was linear in the range of 0.104-20.8 microg (r = 0.9999), the average recovery was 97.68%, RSD 1.4% (n=6).

CONCLUSIONThis HPLC method is simple,accurate and reliable.

Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Eleutherococcus ; chemistry ; Fatigue ; drug therapy ; physiopathology ; Glucosides ; analysis ; Humans ; Male ; Mice ; Phenylpropionates ; analysis ; Swimming

The aim of this study was to investigate effects of IAT and MAT chemotherapeutic regimens treating patients with refractory and relapsed acute myeloid leukemia (AML). 99 patients with refractory and relapsed AML received IAT regimen or MAT regimen as study objects were retrospectively analyzed (56 patients with refractory AML and 43 patients with relapsed AML). Among of them, 28 patients were treated with IAT regimen, and 71 patients received with MAT regimen. The results showed that in 2 groups mentioned above the OR was 65.7%, CR was 49.5%, PR was 16.2%; in IAT group the OR was 64.3%, CR was 46.4%; in MAT group the OR was 66.2%, CR was 50.7%, no statistical difference was found between these 2 groups; The 2 years overall survival was 25% in IAT group and 15.5% in MAT group. Serious infection in IAT and MAT regime groups was 25% and 9.9%, respectively. It is concluded that both IAT and MAT regimens are effective methods for inducing CR in patients with refractory of relapsed AML. IAT and MAT regimens can be used in treatment of the refractory or relapsed MAL patients who were not respond to other regimen.
Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Child ; Cytarabine ; administration & dosage ; Female ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; etiology ; Male ; Middle Aged ; Recurrence ; Retrospective Studies ; Vidarabine ; administration & dosage ; Young Adult

OBJECTIVETo investigate the clinical outcomes of surgical treatment of fracture of the fifth metatarsal base combined with degree III lateral ligament injury of ankle.

METHODSFrom January 2008 to December 2011, 32 patients with fracture of the fifth metatarsal base combined with degree III lateral ligament injury of ankle were treated with surgery. Fractures were fixed with compressed canulated screw and ligaments were repaired with suture anchors. After operation, ankle joints were fixed in neutral position and slightly valgus position by plaster slab. Taking out stitch was performed at 2 weeks after operation and non-weight loading walking by double crutches support started; after the 6 weeks, remove the gypsum and part-weight loading walking by brace protection; at the 8 weeks after operation, completely weight loading walking was permitted. American Orthopaedic Foot and Ankle Society (AOFAS) was used to evaluate the clinical effect.

RESULTSThirty-two patients were followed up from 8 to 18 months with an average of 12 months. All fractures obtained healing with an average time of 12.5 weeks (ranged, 8 to 24 weeks). According to the standard of AOFAS, 18 cases got excellent results and 14 good.

CONCLUSIONThe method that fracture fixation with compressed canulated screw and ligament repair with suture anchors can obtain satisfactory effects in treating fracture of the fifth metatarsal base and degree III lateral ligament injury of ankle.

Adult ; Aged ; Ankle Injuries ; surgery ; Bone Screws ; Collateral Ligaments ; injuries ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Metatarsal Bones ; injuries ; surgery ; Middle Aged

ObjectiveTo explore the surgical techniques and clinical experiences in treating thoracoabdominal aorta dissecting aneurysm following Sun's procedure.MethodsFrom June 2009 to May 2011,thoracoabdominal aorta replacement was performed in thirteen patients with thoracoabdominal aorta dissecting aneurysm following Sun's procedure in Beijing Anzhen Hospital.Among which,eleven were male,and two were female with a mean age 39.3 years (28 -58 years).All cases were Stanford A aortic dissection,and were underwent Sun's Procedure.The pathogeny of the dissection,ten were Marfan's syndrome,and three were hypertension.Thoracoabdominal aorta dissecting aneurysms were all Crawford type Ⅱ,with the diameter( 5.78 ± 1.00 )cm (4.0 -8.0 cm).All the procedures were performed through combined thoracoabdominal incision via the retroperitoneal approach.And thoracoabdominal aorta was replaced by a tetrafurcate graft with short-time interval circulatory arrest.24 - 30 mm tetrafurcate grafts were selected in all patients.The main graft of the tetrafurcate graft was anastomosed to the elephant truck stent graft.Visceral arteries were joined into a patch and were anastomosed to the other end of the main graft.T6 to T12 intercostal arteries were reconstructed by an 8 mm sidearm.Another 8 mm sidearm was anastomosed to the left renal artery.Both 10 mm sidearms were anastomosed to iliac arteries.Among which,seven were underwent by profound hypothermia with circulatory arrest,and six were underwent off pump with normal temperature.ResultsNo early death.Twelve patients were cured and discharged from hospital.Seven patients were underwent profound hypothermia with circulatory arrest.Among which,one patient had paraplegia,sepsis,pseudomembranous colitis,respiratory failure,and died on 102-day after operation.Another one patient had neurological dysfunction and was cured after six-day's dehydrated treatment.The UFCT showed that the tetrafurcate graft was unobstructed,no distortion,no pseudoaneurysms in all patients.The branch to intercostal arteries was obstructed in three cases,but there were no spinal cord ischemia complications.The mean duration of follow-up was (9.6 ±2.1 ) months (range,1 -24 months),with a follow-up rate of 100％.All the tetrafurcate graft were very well,no obstruction,no distortion and no pseudoaneurysms.There were no cerebral,spiral cord and abdominal viscera ischemia complications in all patients.ConclusionThoracoabdominal aorta replacement after Sun's procedure is simple,and it is helpful to reduce complications.Following Sun's procedure,using tetrafurcate graft with short-time interval circulatory arrest and off pump technique in thoracoabdominal aorta replacement is simple,and it is helpful to reduce complications.