In order to establish a method by which the recombinant suicide plasmids integrated on the chromosome could be recircled, A simple method of transconjugative cloning was established with the helper plasmids pMT999 or pRK2013 and fusion strains of Shigella flexneri which were obtained by screening with in vivo expression technology. And the cloning efficiency with this method is very high.

Adolescent ; Adult ; Aged ; China ; epidemiology ; Chronic Disease ; epidemiology ; prevention & control ; Epidemiological Monitoring ; Humans ; Male ; Middle Aged ; Risk Factors ; Young Adult

OBJECTIVETo investigate whether mammalian target of rapamycin (mTOR) kinase was abnormally activated in maldeveloped balloon cells and dysmorphic neurons of focal cortical dysplasia (FCD) with refractory epilepsy.

METHODSA total of 12 archival cases of FCD typeIIwith medically intractable epilepsy treated between 2008 and 2010 were retrieved. Perilesional brain tissue was used as control specimens (n = 8). The expression of phosphorylated p-AKT (Ser473), p-mTOR (Ser2448) and p-P70S6K (Thr389) was investigated by imunocytochemistry.

RESULTSThe expression of p-AKT (Ser473), p-mTOR (Ser2448) and p-P70S6K (Thr389) was found in meldeveloped balloon cells and dysmorphic neurons of FCD. A weak stain in a small amount of pyramid neurons was also found in the control group.

CONCLUSIONAbnormal activation of mTOR in maldeveloped balloon cells and dysmorphic neurons of FCD may be a key molecular mechanism underlying the histological changes and repeated seizures.

Adolescent ; Adult ; Brain Diseases ; metabolism ; pathology ; Child, Preschool ; Epilepsy ; metabolism ; pathology ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Immunohistochemistry ; Male ; Malformations of Cortical Development ; metabolism ; pathology ; Malformations of Cortical Development, Group I ; Nestin ; metabolism ; Neurons ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Ribosomal Protein S6 Kinases, 70-kDa ; metabolism ; TOR Serine-Threonine Kinases ; metabolism ; Young Adult

Objective To explore the effect of treating posterolateral tibial plateau fractures via the fibular osteotomy approach.Methods From August 2009 to August 2011,17 patients with posterolateral tibial plateau fractures,including 12 males and 5 females,aged from 24 to 76 years (average,37.8 years),were treated via the fibular osteotomy approach in our hospital.According to the Schatzker classification,8 cases were type Ⅱ,3 cases were type Ⅲ,6 cases were type V.After operation,X-rays were taken in all patients,and Rasmussen's radiological and functional gradings were used to evaluate radiological and functional outcomes of knees.Results All patients obtained follow-up,ranged from 9 to 35 months (average,18months).The healing time of the fracture ranged from 10.0 to 18.0 weeks (average,13.5 weeks).During the period of followed-up,there was no loss of reduction; one case presented with symptoms of common peroneal nerve injury,such as local hypesthesia in distal lateral lower leg and dorsi pedis,which recovered two weeks postoperatively.According to Rasmussen's radiological grading,the mean score of the knee joint was 17.5(range,14.0 to 18.0).The range of motion of the knee joint ranged from -5°to 135°(average,123.5o).According to Rasmussen's functional grading,the mean score of the knee joint was 26.9 (range,22 to 30).Conclusion Treating posterolateral tibial plateau fractures via the fibular osteotomy approach can obtain sufficient exposure,good reduction and fixation,and avoid flexion contracture of the knee and peripheral vascular nerve injury.Moreover,postoperative function and stability of the knee joint recover well.

Objective To evaluate the clinical outcomes in patients with both column acetabular fractures involvement posterior wall using lag screw through single ilioinguinal approach.Methods Between August 2008 to August 2014,35 consecutive patients with both columns acetabular fractures and fracture also involved posterior wall and fixed by lag screws were retrospectively analyzed.There were 25 males and 10 females.The average age was 44.4+ 12.5 years (range,18-72 years).According to Letournel classifications,the acetabular fracture involved both columns and posterior wall in all cases,and 2 cases with additional seagull sign.The surgeon reduced posterior wall by pressing the fragment through a small tunnel in the soft tissue leaning against the external cortex of iliac bone and fixed the fragment using lag screws from the anterior side.Clinical and functional outcomes were assessed using the modified Merle d'Aubigné scoring system.Radiographic results were evaluated based on Matta scoring system.Results The patients were follow for an average of 44.7+ 18.9 months (range,24-96 months).The average operative time was 257.7±60.4 min (range,160-490 min).The average blood loss during the operating was 742.9±+614.5 ml (range,300-4 000 ml).Loss of reduction of the posterior wall was not found in any case.At the latest follow-up,radiographic results were excellent in 11,good in 19,and poor in 5.The average modified Merle d'Aubigné score was 16.4 (range,13-18).There were 8 cases of excellent,23 cases of good,and 4 cases of fair.Four patients developed deep venous thrombosis of the lower extremities after 3-4 days of operation.Warfarin was used for anticoagulation therapy and thrombi disappeared 6 months postoperatively.One case had superficial wound infection and was treat with vacuum sealed drainage (VSD) and anti-infection therapy.The incision healed without infection after one month.One case with poor quality of reduction and remaining seagull sign was treated with total hip arthroplasty after 35 months because of severe traumatic arthritis.The total reoperation rate was 5.7％ (2/35).latrogenic sciatic nerve injury as well as heterotopic ossification was not found in any case.Conclusion Satisfactory quality of reduction and good clinical outcomes can be achieved in patients with acetabular fractures involved both columns and posterior wall by single ilioinguinal approach and lag screw fixation for posterior wall.Iatrogenic sciatic nerve injury as well as heterotopic ossification was not found in any case.

Objective To evaluate the clinical outcomes of ilioinguinal approach with short infrapectineal buttress plate fixing posterior column for patients with complex acetabular fractures. Methods Data of 29 consecutive patients (male 22, fe?male 7, age range 25-72 years, average age 53 ± 6.3 years) with complex acetabular who had operated by single ilioinguinal ap?proach with infrapectineal buttress plate from September 2008 to August 2012 were retrospectively analyzed. According to Letour?nel?Judet classifications, there were 4 cases of anterior column and posterior hemi?transverse, 11 cases of associated both?column, 4 of T?shaped and 6 with seagull sign. The acetabular fractures model was printed preoperatively by 3D printing technique for sur?gical simulation, open reduction and internal fixation through single ilioinguinal approach with a short infrapectineal plate fixing posterior column, particularly an ox horn shape K?wire sleeve was developed for drilling and screw insertion using flexible screw?driver. Clinical, radiographic, and functional outcomes assessed by the modified Merle d’Aubigné score were collected. Results Two patients were lost to follow?up, including one patient who died at 15 days postoperative because of pulmonary embolism, and the other one who had moved abroad at 12 months postoperative. The remaining 27 (93%) had a mean follow?up of 40 months (range, 24-75 months). The operating time was 180±35.5 min;the time for the fracture union was 3.5±0.9 month;blood loss during the operating was 500±43.9 ml;no case had fracture re?displacement. At the latest follow?up, radiographic grades were excellent in 17, good in 8, poor in 4, including one poor patient who had a total hip arthroplasty (3%) at 35 months after the internal fixation. The average modified Merle d’Aubigné score was 16 (range, 10-18), and categorized as excellent in 12, good in 8, fair in 5, and poor in 2. Three patients had developed deep venous thrombosis of the lower extremities, and one of them died of pulmonary embo?lism 15 days after operation; two cases of deep venous thrombosis of the lower extremity were found 3-4 days after operation. Thrombosis disappeared 6 months after anticoagulant therapy by warfarin. One case who had superficial wound infection after oper?ation was treated by anti?infection and VSD negative pressure, and incision was healed after one month. One case with poor quality of fracture reduction and seagull sign was treated with total hip arthroplasty after 35 months because of traumatic arthritis. The to?tal complication rate was 24%(7/29). Conclusion In the patient with complex acetabular fractures combining posterior column medial displacement, single Ilioinguinal approach with infrapectineal buttress plates could achieve a stable concentric hip joint, and immediate postoperative ambulation can result in reasonable clinical, radiographic, and functional outcomes.

OBJECTIVETo elucidate the clinicopathological features of solid pseudopapillary tumor (SPT) of the pancreas.

METHODSEight patients with SPT of the pancreas admitted from August 1996 to March 2005 were retrospectively analyzed.

RESULTSAll the 8 patients were female with an average age of 25.3 (13-41) years. The primary clinical manifestations included abdominal mass (n = 3), vague abdominal pain (n = 3), and duodenal obstruction (n = 1). SPT was occasionally found in one patient during physical examination. Six tumors located at the head and the other two in the body and tail of the pancreas. Pancreaticoduodenectomies were performed in 4 patients, tumor enucleations in 2, distal pancreatectomies in 1, and palliative internal drainage with a cystoenterotomy in the other one with an unresectable huge cystic lesion. All patients were alive on an average follow-up of 37.8 (8-103) months.

CONCLUSIONSPT occurs mainly in adolescent and young females, and satisfactory outcome may be achieved with active and appropriate surgeries.

Adolescent ; Adult ; Carcinoma, Papillary ; diagnosis ; pathology ; Female ; Humans ; Pancreas ; pathology ; Pancreatic Neoplasms ; diagnosis ; pathology ; Prognosis

OBJECTIVETo investigate the relationship between chromobox protein homolog 7 (cbx7) expression and the occurrence and development of colorectal carcinoma (CRC), gastric carcinoma (GC) and hepatocarcinoma (HCC) tissues.

METHODSThe samples of neoplastic tissues and the corresponding cutting-edge normal tissues from 22 cases of CRC, 20 cases of GC, 30 cases of HCC were surgically collected. Level of cbx7 mRNA was detected with a fluorescent quantitative RT-PCR assay, and the correlationship among expression of cbx7 mRNA, the patients' clinicopathologic features and the surviving time after surgery was analyzed.

RESULTSThe relative copy number of cbx7 mRNA in carcinomas and the normal tissues was 0.010 ± 0.015 vs 0.053 ± 0.042 for CRCs, 0.197 ± 0.195 vs 1.891 ± 1.254 for GCs, and 0.008 ± 0.008 vs 0.030 ± 0.021 for HCCs, respectively. Compared with the corresponding normal tissues, cbx7 expression was significantly downregulated in CRCs, GCs, and HCCs (t = -7.351, -5.417 and -6.680, respectively, P < 0.01). The expression of cbx7 mRNA in CRCs had significant differences not only between two age groups (the relative copy number of cbx7 mRNA in age > 55 group was 0.007 ± 0.015, but 0.017 ± 0.012 in age ≤ 55 group, t = -2.586, P = 0.022); but also between vascular embolus-positive and negative groups (the level of cbx7 mRNA in positive and negative group was 0.022 ± 0.021 vs 0.006 ± 0.011, t = -3.175, P = 0.010). The area under the receiver operating characteristics (ROC) curve is 0.769 (P = 0.033). when the Cut-off value of the relative copy number of cbx7 mRNA was 0.002 in CRCs. The values less-than 0.002 were defined as low expression. The CRC patients with low expression of cbx7 had a shorter overall survival time; whose 5 years survival rate was only 30.8% (4/13); while the rate was 77.8% (7/9) in high expression of cbx7 group. The difference had statistical significance (χ(2) = 4.329, P = 0.037). The similar differences could not be found among GC and HCC patients.

CONCLUSIONDownregulation of cbx7 expression was very common among multiple carcinomas cases, and the downregulation influenced the prognosis of CRC patients.

Colorectal Neoplasms ; genetics ; metabolism ; Down-Regulation ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms ; genetics ; metabolism ; Male ; Middle Aged ; Neoplasms ; Polycomb Repressive Complex 1 ; Repressor Proteins ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism

Objective To assess the effect of preoperative administration of tranexamic acid (TXA) on the hidden blood loss in the surgery of intertrochanteric fractures with proximal femoral nail anti-rotation (PFNA).Methods Eighty patients with intertrochanteric fracture were treated with PFNA in our hospital from November 2015 to July 2016.They were 15 men and 65 women,with a mean age of 72.6 years.Of them,39 were included into TXA group where TXA was administered preoperatively and 41 were assigned into the control group where no TXA was used preoperatively.Blood routine examinations were carried out on one day before operation,the first and third days after operation.The surgical blood loss,operative blood transfusion,24-hour drainage after operation,and postoperative 3-day blood transfusion were recorded.The total and hidden blood losses were calculated according to the formula.The 2 groups were compared in terms of blood loss and complications.Results In TXA group,the total blood loss (1,632.3 ± 849.2 mL),the hidden blood loss (1,270.9 ± 623.3 mL) and the transfusion rate (28.2％) were significantly lower than those in the control group (2,014.8 ± 924.7 mL,1,549.1 ± 624.9 mL and 56.1％) (P ＜ 0.05).There was no significant difference between the 2 groups in visible blood loss (361.4 ± 154.3 mL for TXA group versus 465.7 ± 191.3 mL for the control group) (P ＞ 0.05).Deep venous thrombosis occurred in 2 patients,limb swelling in one patient and wound dehiscence in one patient in TXA group while deep venous thrombosis occurred in one patient,limb swelling in 3 patients and wound dehiscence in none in the control group,showing no significant differences between the 2 groups (P ＞ 0.05).No infection was observed in either group.Conclusion Preoperative administration of TXA can reduce surgical hidden blood loss and transfusion rate as well but will not increase the risk of deep vein thrombosis in the surgery of intertrochanteric fractures with PFNA.