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

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

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

OBJECTIVEPancreatic metastasis from renal cell carcinoma (RCC) is a rare event and has not been reported in our country. We report a series of 3 patients with metastatic RCC to the pancreas after radical nephrectomy at our institution. The published reports in the literature were reviewed, and the diagnosis, treatment as well as prognosis of this rare event were discussed.

METHODSThe data of 3 RCC patients with metastasis to the pancreas were reviewed retrospectively, including radical nephrectomy, metastatic interval, the second and third surgical removal. Survival of the three patients was analyzed and the reports in the literature were compared as well.

RESULTSThe average interval from radical nephrectectomy to the comfirmed pancreatic metastasis was 6.6 years (range, 1.2 to 12 years). The pathological stage revealed T2N0M0 (n = 2) or T3N0M0 (n = 1), with right-sided tumor in 2 patients and left side in 1. One patient was asymptomatic, while the other two cases were symptomatic at presentation, including upper abdominal pain, weight loss, slight xanthochromia of the skin and titillation, clay stool (n = 1); irregular fever, weight loss and jaundice (n = 1). All pancreatic metastases were hypervascular on arterial stage of CT imaging. One patient had only a solitary pancreatic metastasis (n = 1), the another showed two metastatic lesions (n = 1), the third one had multiple lesions (n = 1). Surgical removal was accomplished in 2 patients: including pylorus-preserving pancreaticoduodenectomy in one, and pylorus-preserving pancreaticoduodenectomy together with partial tail resection in another one. The third one only received interventional therapy due to widespread extrapancreatic metastasis, and died of disseminated disease 11 months after the therapy. One of the above two surgically treated patients underwent the second removal due to local recurrence 2.5 years after the first removal of pancreatic metastasis. These two patients were still alive after follow-up of 8.6 years and 16.1 years, respectively.

CONCLUSIONRenal cell carcinoma is an unpredictable tumor that may demonstrate very delayed metastasis even from early-stage of the disease. The pancreas is a rare site of metastasis from renal cell carcinoma. We advocate careful long-term follow-up of patients with a history of RCC. Aggressive surgical management of pancreatic metastatic lesions may provide a chance of long-term survival.

Aged ; Carcinoma, Renal Cell ; pathology ; surgery ; Chemotherapy, Cancer, Regional Perfusion ; Female ; Follow-Up Studies ; Humans ; Kidney Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Nephrectomy ; methods ; Pancreatic Neoplasms ; diagnosis ; drug therapy ; secondary ; surgery ; Pancreaticoduodenectomy ; methods ; Retrospective Studies ; Tomography, X-Ray Computed ; Ultrasonography, Doppler, Color

Inherited afibrinogenemia is a rare autosomal recessive bleeding disease characterized by complete absence of fibrinogen in blood. To identify the genotype in a Chinese family with inherited afibrinogenemia, the samples of peripheral blood were collected from 6 members of 3 generations. The activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT) and fibrinogen (Fg, clauss) were tested. Fg was also analyzed by using immunoturbidimetry method. DNAs of six members were extracted by using a DNA extract kit. All the exons and exon-intron boundaries of the three fibrinogen genes were amplified by using PCR and analyzed by direct sequencing. The results showed that the parents of proband were 3 degree consanguinity. A homozygous c.934_935insA in FGA was found in proband which results in the change of protein p.Ser312fsX42. The parents, grandmother, maternal grandmother and father's sister were all detected with heterozygous mutation which was same as that in proband. In conclusion homozygous c.934_935insA in FGA is a cause of inherited afibrinogenemia and a novel mutation being reported.
Afibrinogenemia ; etiology ; genetics ; Child ; Exons ; Female ; Fibrinogen ; genetics ; Frameshift Mutation ; Heterozygote ; Humans ; Male ; Pedigree

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