Objective To investigate the role of autophagy in radiation-induced death process of human esophageal squamous carcinoma Eca-109 cells.Methods Esophageal carcinoma cell line Eca-109 was divided into 6 groups of control,5 mmol/L 3-Methyladenine treatment,10 mmol/L treatment,6 Gy irradiation,irradiation + 5 mmol/L drug,and irradiation + 10 mmol/L drug.Some cells were transferred with GFP-LC3 plasmid and the changes of autophagosome were obserred.After each treatment,the expression of autophagy marker LC3B was measured by Western Blot,cell viability was detected by MTT,morphological characteristics of apoptosis cells were stained with a fluorescein of Hoechst 33342 and the percentage of apoptotic cells and cell cycle distribution were measured by flow cytometry.Clonogenic survival were used to evaluate the cell radiosensitivity.Results Autophagy level was increased after radiation,and the LC3B Ⅱ expression and LC3B Ⅱ/LC3B Ⅰ ratio were significantly decreased by autophagy inhibitor 3-Methyladenine (F =25.64,P ＜ 0.05).The number of autophagosome fluorescent foci were significantly increased in the GFP-LC3 transfected cells after radiation,but reduced by 3-Methyladenine (F =127.36,P ＜ 0.05).Compared with radiation alone group,autophagy inhibition combined with radiation significantly decreased cell viability (F =129.54,P ＜ 0.05) and colony formation,increased apoptosis and the percentage of G2/M-phase cells.Conclusions 3-Methyladenine enhances the radiosensitivity of esophageal squamous carcinoma Eca-109 cells,suggesting that inhibition of autophagy could be used as an adjuvant treatment of radiotherapy in esophageal squamous carcinoma.

Objective To improve the effect of clinical diagnosis and treatment of acetabular fracture with a femoral nerve injury by analyzing the causes of femoral nerve injury following acetabular fractures.Methods From January 1996 to November 2004,146 cases of acetabular fractures were treated operatively.Six cases of them were complicated with femoral nerve injury.The causes of femoral nerve injury were analyzed on the basis of clinical manifestations,CT scan and 3-dimensional reconstruc- tion.All the cases were classified according to Letournel and Judet classification.Three cases had hema- toma compression (2 cases with double column fractures and 1 with transverse-posterior wall fractures);2 cases had femoral nerve injury caused by fracture fragments (1 case with anterior wall fracture following anterior hip dislocation,the another with old fracture of anterior column combined with fracture of superior ramus of pubis);and one case had anterior column fracture combined with fracture of wing of ilium,and the femoral nerve was injured by traction in operation.Clearance of hematoma,nerve tract decompression and epineuria solution were performed in 5 cases,and 1 case was treated conservatively.Results The average follow-up period was 1.8 years(ranging from 1 to 3 years).The muscle power of quadriceps fem- oris recovered from 1-2 grade before operation to 4-5 grade after operation in 5 cases.The function of ex- tensor knee and gait was normal.The function of sensory completely recovered in 4 cases.One case was followed up for 2 years,which showed the patient still suffered from hypoesthesia in the lower 2/3 of the thigh and the medial of the leg.One ease of traction injury was followed up for 1.2 years,showing the muscle power recovered to normal,but still presented with sensory disability.Conclusion Acetabular fractures associated with femoral nerve injury are rare.For complex acetabular fractures and severe trau- ma,attention should be given to the possibility of femoral nerve injury.Fragment stabbing and compres- sion of hematoma around iliopsoas muscle are the common causes of femoral nerve injury following ace- tabular fractures.Iatrogenic injury should not be ignored.

OBJECTIVETo investigate the effects of basic fibroblast growth factor (bFGF) on the expression of glial fibrillary acidic protein (GFAP) after tractive spinal cord injury in rats and to explore the recovery of spinal cord function.

METHODSThe rats were subjected to tractive spinal cord injury at T13-L2. Cortical somatosensory-evoked potential (CSEP) was closely monitored and when P1-N1 wave amplitude decreased to 70% of that before operation, a small-bore catheter was inserted below the injured plane through subarachnoid cavity. In the treatment groups, 20 microl of bFGF solution (containing 20 microg of bFGF) was injected through the catheter right after the operation and 1, 2, 3, 4, 8, 12 and 24 h postoperatively. In the control group, same volume of normal saline was injected and every four rats were killed at 1, 4, 7, 14 and 21 d after the operation. Combined behavior score (CBS) and electro-physiological examination were adopted to evaluate function recovery. Expression of GFAP was observed by immuno-histochemical staining and was analyzed quantitatively by computer image analysis.

RESULTSThere was statistically significant difference in GFAP-positive cells between bFGF treatment group and the control group (P<0.01). Similar tendency was indicated by the results of CBS and CSEP.

CONCLUSIONSbFGF can induce large expression of GFAP after tractive spinal cord injury in rats and promote spinal function recovery, which is highly important for spinal cord regeneration.

Animals ; Disease Models, Animal ; Evoked Potentials, Somatosensory ; drug effects ; Fibroblast Growth Factor 2 ; pharmacology ; Glial Fibrillary Acidic Protein ; drug effects ; metabolism ; Immunohistochemistry ; Rats ; Rats, Sprague-Dawley ; Recovery of Function ; Reference Values ; Spinal Cord Injuries ; metabolism ; physiopathology ; Traction

OBJECTIVETo establish an animal model of tractive spinal cord injury in rats in order to investigate its pathophysiological changes and clinical significance.

METHODST(12)-L(3) spines were tracted longitudinally with a special spinal retractor that was put on the proccessus transverses of T(12)-L(3) vertebrae of the rat after exposing T(13)-L(2) spinal cord via dual laminectomy. At the same tine, the spinal cord function was monitored by cortical somatosensory evoked potential (CSEP). Rats were randomly divided into four groups according to the amplitude of CSEP P(1)-N(1) wave, the amount of the decreasing P(1)-N(1) wave was 30% (the 30% group), 50% (the 50% group) and 70% (the 70% group), respectively. After traction, the changes of the neural behavioral function in rats were observed and the morphological structure of the spinal cord was analyzed quantitatively with image analysis system of computer.

RESULTSWith traction of spine, compared with the control group, the 30% group had no marked difference in combined behavioral score (CBS), neuron count, section area of neuron and Nissl body density, but the 50% and 70% groups had marked difference (P<0.01). Light microscope showed that the neuron volume was slightly small and the Nissl body was reduced lightly in the 30% group; the neuron space was enlarged and the neuron was degenerative, reductive, and dissolved, and the spinal cord structure was destroyed in the 50% and 70% groups.

CONCLUSIONSThe animal model of tractive spinal cord injury in rats is a reproducible, graded and clinic mimic. The model in this article provides a valuable assistance in further understanding etiopathology and screening effective measures of therapy and prophylaxis of the injury.

Animals ; Female ; Male ; Models, Animal ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; physiopathology ; Traction

Adult ; Hemophilia A ; complications ; Humans ; Leukemia, Promyelocytic, Acute ; complications ; Male

OBJECTIVETo observe and evaluate the clinical effects of surgical treatment for the accessory navicular syndrome with simple excision.

METHODSFrom November 2006 to December 2008, 23 patients (twenty-five feet) with accessory navicular syndrome received simple excision of the accessory navicular bone. The chief complains were intermittent pain of feet after running or walking. Physical examination showed local tenderness on palpation in the region of the navicular bone. X-ray or CT showed there was an accessory navicular bone. The present history ranged from 6 months to 12 years. There were 14 males and 9 females. The mean age was 14.6 years, ranging from 8 to 35 years. About 2 cm long incision was made at the tip of the medial prominence of the navicular bone. After partial dissection of the posterior tibial tendon, the accesssory navicular bone was exposed and excisied. The prominence of the tuberosity of the navicular bone was cut and shaved. The posterior tibial tenden was repaired before closing the wound. The foot was immobilized with cast or brace in inversion position and no weight-bearing for 2 weeks. Strenuous jumping or dancing must be avoided in 3 months after surgery. The patients with residual symptoms and signs received physical therapy and an arch support for shoes without flatfoot deformity.

RESULTSThe average clinical follow-up during was 12 month (ranged, 3 to 18 months). The excellent results in 21 feet and good in 4 feet (3 feet with mild flat deformity and 1 foot with old sprain injury). The average hospital stay was 5 days and no wound infection occurred. All patients resumed the normal life and study after operation.

CONCLUSIONSurgical treatment of the accessory navicular syndrome with simple excision has the advantages of less invasive to the posterior tibial tenden and the medial longitudinal arch of the foot, shorter time of immobilization of the foot and stay in hospital, small incision and good clinical results. This procedure is one of the best selective treatments for the accessory navicular syndrome, especially for the patients without flatfoot deformity and old sprain injury.

Adolescent ; Adult ; Bone Diseases ; pathology ; surgery ; Child ; Female ; Foot Diseases ; pathology ; surgery ; Humans ; Male ; Young Adult

OBJECTIVETo study the expression of specific anti- platelet glycoprotein autoantibodies GP II b/III a, GP I b/IX and GP I a/II a in primary immune thrombocytopenia (ITP), and to evaluate the relationship between the therapeutic effect and the expression of specific anti- platelet glycoprotein antibodies GPIIb/IIIa, GPIb/IX and GPIa/IIa.

METHODSAnti-GPIIb/IIIa, GPIb/ IX and GP I a/II a antibodies were assayed by ELISA for patients with ITP. Total 442 patients in our hospital, who were retrospectively investigated from December 2010 to November 2012, were divided into newly diagnosed ITP, persistent and chronic ITP. The expression of specific anti- platelet glycoprotein antibody in each group was measured separately. The newly diagnosed ITP patients were treated with intravenous IgG (IVIG) and corticosteroids. The relationship between the expression of specific anti- platelet glycoprotein antibodies GPIIb/IIIa, GPIb/IX and GPIa/IIa and the complete response (CR) was studied.

RESULTSPositive rates of anti- platelet glycoprotein antibodies were 59.09%, 26.97% and 37.35% respectively in newly diagnosed ITP, persistent and chronic ITP, the difference was statistical significant (P<0.05). In newly diagnosed ITP, positive rate of antibody against GPIIb/IIIa was 38.64%, double positive rate of antibodies against both GP II b/III a and GP I a/II a was 15.91%, there was statistical significance (P<0.05) compared with that of persistent and chronic ITP. The complete response (CR) rate in newly diagnosed ITP patients with positive antibody against GP II b/III a was 80.39% after treatment with IVIG and corticosteroids. There was statistical significance compared with that in patients having no antibodies (P<0.05).

CONCLUSIONThe expression of antibodies against GP II b/III a and double positive for both GP II b/III a and GP I a/II a autoantibodies increased in newly diagnosed ITP patients. Patients with anti-GP II b/III a autoantibody had good response to medication with IVIG and corticosteroids.

Adolescent ; Adult ; Aged ; Autoantibodies ; metabolism ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Platelet Glycoprotein GPIIb-IIIa Complex ; immunology ; Platelet Glycoprotein GPIb-IX Complex ; immunology ; Platelet Membrane Glycoproteins ; immunology ; Retrospective Studies ; Thrombocytopenia ; drug therapy ; immunology ; metabolism ; Treatment Outcome ; Young Adult

OBJECTIVETo evaluate the expression of TEIF protein in human tumors of soft tissue and its significance.

METHODSAnti-TEIF polyclonal antibody was generated by immunization of E.coli expressed His-TEIF protein. The expression of TEIF in 166 cases of sarcomas and 28 case benign tumors or tumor-like lesions of soft tissue arranged in tissue chip was analyzed by immunohistochemistry.

RESULTSPolyclonal antibody obtained from immunized rabbit was verified in Western blot to prove its specific reactivity with native TEIF protein. The immunohistochemical staining of TEIF showed that about 58% (97/166) of sarcomas were positive and significantly different from that of benign tumors or tumor-like lesions (11%, 3/28). The positive staining was predominantly in synovial sarcoma 94% (16/17), primitive neuroectodermal tumor (PNET) 91% (21/23), both of which were significantly higher than 43% (6/14) of dermatofibrosarcoma protuberans, 38% (6/16) of myxofibrosarcoma, 36% (8/22) of malignant peripheral nerve sheath tumor, 32% (6/19) of liposarcoma, (P < 0.05, respectively), but not higher than 75% (15/20) of malignant fibrous histiocytoma, 70% (7/10) of rhabdomyosarcoma or 64% (9/14) of leiomyosarcoma. Meanwhile, strong positive staining of TEIF (>or= 2+) was frequently observed in PNET (83%, 19/23) and synovial sarcoma (76%, 13/17). With respect to FNCLCC grading, 19 cases of grade I sarcoma TEIF was 32% (6/19) and strong positive was 11% (2/19), 44 cases of grade II sarcoma was 48% (21/44) and 32% (14/44), and 70 of grade III was 84% (59/70) and 70% (49/70). The rate of either positive or strong positive in grade III sarcoma was significantly different from that of either grade I or II (P < 0.05), but no difference between the latter two groups (P > 0.05).

CONCLUSIONSTEIF protein could be detected in large part of soft tissue sarcomas, and it not only over-expressed in most of PNET, synovial sarcomas, but also correlated with histological grading.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Cell Line, Tumor ; Child ; Child, Preschool ; Female ; HeLa Cells ; Histiocytoma, Malignant Fibrous ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Infant ; Leiomyoma ; metabolism ; pathology ; Male ; Middle Aged ; Neuroectodermal Tumors, Primitive ; metabolism ; pathology ; Rhabdomyosarcoma ; metabolism ; pathology ; Sarcoma ; metabolism ; pathology ; Sarcoma, Synovial ; metabolism ; pathology ; Soft Tissue Neoplasms ; metabolism ; pathology ; Tissue Array Analysis ; Transcription Factors ; biosynthesis ; Young Adult

OBJECTIVETo investigate whether the plasmid bearing attB and human coagulation factor IX (hFIX) coding sequence could insert into hemophilia B mice genome and persistently express hFIX with co-injected integrase.

METHODSThe plasmid attB-hFIX-pIRES2-EGFP was constructed, which bore attB site and hFIX coding sequence and was proved in vitro to express hFIX. The plasmid and CMV-int expressing integrase was co-infused rapidly in a large-volume solution through tail vein of hemophilia B mice. Mice infused with the plasmid alone served as controls. ELISA was performed to determine serum hFIX level. Correction of coagulation defect in vivo by plasmid infusion was assessed by bleeding time. Genomic integration of the plasmid was determined by nested PCR.

RESULTSThe plasmid attB-hFIX-pIRES2-EGFP was successfully constructed. The hemophilia B mice produced (1533 ± 239) ng/ml hFIX at 24 hour after infusion of the hFIX encoding plasmid and the bleeding diathesis of the hemophilia B mice was significantly corrected as measured by clotting assays. However, whether or not co-injected with CMV-int, the serum hFIX level decreased to background level in 10 days after infusion. Nested-PCR results indicated that the integrase phiC31 resulted in the integration of the plasmid in the mouse liver chromosomes.

CONCLUSIONIntegrase phiC31 can catalyze recombination of 34 bp attB and pseudo-attP. Human FIX driven by CMV promoter can be transiently and highly expressed after infusion, but rapidly silenced in vivo.

Animals ; Factor IX ; genetics ; Gene Expression ; Genetic Therapy ; Genetic Vectors ; Genomics ; Hemophilia B ; therapy ; Humans ; Hydrodynamics ; Mice