Phage display technology is improved in recent years.LoxP-cre system based in vivo recombination is the most important one. From double recombination system to single recombination system,from large scale sample sequencing to visual judgement, in vivo recombination makes it feasible to construct large natural antibody library, and it is favorable to get high affinity antibody. This review introduces the important improvement of in vivo recombination system.

Objective To assess the clinical walue of 3D retroperitoneal laparoscopic renal pedicle disconnection by comparing its clinical effects with those of traditional 2D laparoscopic.Methods Clinical data about 49 cases from our department with chyluria respectively treated by 3D retroperitoneal laparoscopic and 2D laparoscopic renal pedicle disconnection were analyzed retrospectively.They were divided into 3D group(26 cases)and 2D group(23 cases),and operated by the same salty doctor.The operation time,intraoperative blood loss,postoperation hospital stay and recurrence rate of the two group were compared.Results The operation time in 3D group[(95.2±30.3)min] was shorter than 2D group[(120.4±25.8)min,P<0.05].The intraoperative blood loss in 3D group[(30.0±12.7)ml] was less than 2D group[(55.0±21.7)ml,P<0.05].There were no difference in postoperation hospital stay between the two groups.All of the cases were not recurred.Conclusion 3D Retroperitoneoscopic renal pedicle lymphatic disconnection for treatment of chyluria is safty and feasible.3D laparoscopy provides high-quality 3D endoscopic view and facilitates precise manipulation during surgery,resulting in shorter operation time and intraoperative blood loss compared with 2D laparoscopy.

?Objective:To analyze the therapeutic characteristic of mandibular prognathism by means of orthognathic surgery and orthodontic treatment.Methods:125 patients with skeletal mandibular prognathism were treated by intraoral ramus osteotomy or mandibular body osteotomy,They also accepted different preoperative and postoperative orthodoitic treatment according to different surgery.Results: Satisfied results were obtained through combined treatment of orthognathic surgery and orthodontics.The effective rate of intraoral ramus and mandibular body osteotomy was 88.3% and 83.3% respectively .Two patints treated with mandibular body osteotomy relapsed in two years,and mandibular body osteotomy was performed again to correct the diformity.Conclusion:proper pre and postoperative orthodontic treatments are necessary in order to assure asethetics effect and ideal occlusion in new position.

Objective:To study Clinical Effect of Anterolateral Approach ofor the Patients with Hip Arthroplasty and the effect on the serum CRP,IL-6,D-Dimer in levels.Methods:102 Patients with Hip Arthroplasty who received therapy from February 2013 to January 2015 in our hospital,Patients were asked to be divided into observation group and control group.Among them,the control group of 54 cases,through the conventional approach to hip arthroplasty;48 cases of observation group.Serum CRP,IL-6 and D-dimer levels were compared between the two groups before and after surgery,and the efficacy of the two groups was compared by follow-up.Results:After treatment,the total effective rate of the observation group was significantly higher than that of the control group (P＜0.05),Serum CRP,IL-6 and D-dimers increased in both groups at 12 h after surgery and decreased gradually at 7 and 14 days.The levels of serum CRP,IL-6 and D-dimer in the observation group were significantly lower than those in the control group at 12 h,7 d and 14d(P＜0.05).Conclusion:The clinical effect of the anterolateral approach on hip arthroplasty is more significant than that of conventional approach,and can effectively reduce the levels of serum CRP,IL-6 and D-dimer,and is more favorable for postoperative recovery.

Objective To study the long term effect and practical value of unilateral cleft lip repair with extended Mohler surgery based on over 10 years experiences.Methods A lot of 320 cases with unilateral cleft lip were repaired with extended Mohler surgery at average age of 5 months and 20 days.65 repaired cases were followed up and long term treatment result was evaluated.26 cases were evaluated in 10.5 to 15 years after surgery.39 cases were evaluated in 6 to 24 months after surgery.And the details of surgery were described.Results Excellent results for most of cleft patients (63 cases, 97%) were achieved, which included unobvious vertical scar symmetric to unaffected philtrum column, symmetric cupid's bow, nostrils and intermedial nasal columellae.Conclusions Extended Mohler surgery is great valuable for unilateral cleft lip repair.Appropriate indications should be stressed.

Objective To explore the ultrasound imaging and the clinical analysis of eosinophilic cystitis in children and to improve the understanding of this very rare clinical disease.Methods The ultrasound imaging of 9 patients who were diagnosed as eosinophilic cystitis in Nanjing Children's Hospital Affiliated to Nanjing Medical University from Jul.2007 to May 2013 were analyzed retrospectively,and the relevant literatures were reviewed.Results The ultrasound imaging of the 9 patients were described as follows:4 cases showed a diffuse thickening bladder wall,2 cases showed a localized mass-like thickening bladder wall,and 3 cases showed a markedly thickening anterior bladder wall and bilateral bladder wall.Laboratory studies showed that all the patients were presented with eosinophilia.Six patients underwent cystoscopy that showed an erythematous,and velvety appearance of the bladder mucosa.Histopathology disclosed a bladder mucosal tissue with dense interstitial eosinophilic infiltration consistent with eosinophilic cystitis.Three patients had clinical resolution under the treatment of anti-infections and urine alkalinization.Other 6 patients received prednisone at 0.5-1.0 mg/kg and an antihistamine treatment.Prednisone was eventually tapered off after 2-week reemission.Regular follow-up by urinary bladder ultrasound and urine routine did not show any disease recurrence in 9 children.Conclusions The combination of bladder wall thickening and peripheral eosinophilia may suggest the likely diagnosis of eosinophilic cystitis.Cystoscopy with bladder biopsy is the most important step in the diagnosis of eosinophilic cystitis.Anti-histamines and corticosteroids can be used as the primary management and have good results.

Objective To explore the role of severity of limb ischemic injury in the treatment of popliteal artery injury.Methods A retrospective analysis was conducted of 259 patients with popliteal artery injury who had been treated from January 2002 to December 2014.They were 187 males and 72 females with a median age of 37.6 years (range,from 14 to 68 years).The time from injury to revascularization was from 4 hours to 29 days.According to the Rutherford classification system,the limb ischemic injury was categorized into 4 grades:grade Ⅰ:presence of sense and motion;grade Ⅱ:presence of sense but loss of active motion;grade m:loss of sense or motion;grade Ⅳ:stiffness.For all the patients with injury of grades Ⅰ and Ⅱ,limb salvage was adopted and opening decompression to osteofascial compartment was performed depending on the intraoperative and postoperative limb swelling.For the patients with grade Ⅲ injury,opening decompression to osteofascial compartment was performed to explore the activity of muscles in those with a strong will to preserve their limbs.When the muscle activity was poor but bright red bleeding was found at the broken muscle ends and there were no fish-like changes in muscles,limb salvage was suggested;otherwise amputation was chosen.For all the patients with grade Ⅳ injury,amputation was suggested.Results Of the 146 cases of limb salvages,142 were successful and 4 failed due to uncontrollable infection,giving a total success rate of 54.83％ (142/259).Complete rupture of the popliteal artery occurred in 23 cases,incomplete rupture or tear in 17,contusion and thrombosis in 219.No patient died due to popliteal artery injury or its complications.The patients with injury of grades Ⅰ,Ⅱ,Ⅲ and Ⅳ were respectively 51,88,67 and 53;their success rates of limb salvage were respectively 100.00％ (51/51),96.59％ (85/88),8.96％ (6/ 67) and 0.For the patients whose time from injury to revascularization was ＜ 6 h,6 h to 12 h,13 h to 24 h,25 h to 1 w,and ＞ 1 w,the success rates of limb salvage were respectively 80.77％ (42/52),54.58％ (73/134),20.51％ (8/39),33.33％ (6/18),and 81.25％ (13/16).The 142 patients whose limbs had been salvaged obtained an average follow-up of 31.4 months (range,from 6 months to 8 years).Of them,those with grade Ⅰ injury obtained normal plantar sensation and active ankle flexion and extension.Of the 85 patients with grade Ⅱ injury,all recovered normal or nearly normal plantar sensation,29 achieved partial recovery of active ankle and digital flexion and extension,but the other 56 obtained no recovery of active ankle motion.Of the 6 patients with grade l injury,none obtained recovery of motion function,4 achieved partial recovery of plantar sensation but the other 2 had no recovery.Conclusion The classification of popliteal artery injury into 4 grades according to its severity of ischemic injury can provide helpful guidance to the treatment and prognosis assessment of the popliteal artery injury.

Objective To investigate the expression and clinical significance of H1 receptor in kidney and bladder tissue of rats after long term ketamine intraperitoneal injection.Methods This study was conducted from May 2012 to December 2012.Sixty male 2-month-old SD rats,weighted (200±10) g,were randomly divided into Group A and Group B.Each group concluded 30 rats.In the Group A,Ketamine (100 mg/kg) was given as intraperitoneal injection every other day,while normal saline (100 mg/kg) was given in Group B.The dosage was adjusted every week according to the weight of rats.After 2,4 and 6 months,10 rats from each group were randomly chosen.First,the micturition number during 2 h was recorded.Then,urine samples over a 24 h period were collected and the content of Na+ and K+ were determined.Finally,the blood samples were obtained from the apex of heart for the creatinine determination.The kidneys and bladders were harvested after the rats were sacrificed.HE staining was conducted on all the tissues.Immunohistochemical S-P method was used to detect the expression of H1 receptor in the bladder and kidney tissues from Group A and Group B.The average optical density (A Value) in each group was separately calculated by Imagepro-plus 6.0 software.All the parameters,mentioned above,were carefully compared.Results The successive rate of establishing rats model was 90％ (9/10),according to the pathological result after 6 months injection.The urine volume of 24 h in group A and B were (15.9±1.3) and (10.1±0.8) ml,respectively.Micturition frequency during 2 hours in group A and B were (6.9±1.4) and (3.0±0.5) times.The urine volume of 24 h and micturition frequency during 2 hours were significantly increased in group A (P＜ 0.05).The urine sodium within 24 h in group A was (1.7±0.1) mmol,which is increased significantly than that in group B (1.0±0.1 mmol).While the urine potassium was less in group A (1.1±0.1 mmol/d) than in group B (2.6±0.1 mmol/d) (P＜0.05).But the serum creatinine level were (60.5±6.8) and (58.1± 3.9) μmol/L in group A and B,which had no difference between the two groups (P＜0.05).The expression of H1 receptor in kidneys and bladder in group A was significantly raised compared with group B (P＜0.05).In the group A,the expression of H1 receptor level in kidney was 0.008±0.001,0.016±0.001,0.023±0.004 after 2,4 and 6 months drug used.The expression level in group A were significantly difference than that in group B (0.003±0.001,0.004±0.002,0.003±0.001) (P＜0.05) and goes up with prolonging the drug using.While in the bladder tissue,the level of H1 receptor expression was 0.017±0.006,0.031±0.012,0.036±0.007 in group A and 0.015±0.007,0.016±0.005,0.016±0.004 in group B,which could be noticed a significantly increasing in group A (P＜0.05).In 4 and 6 months,the H1 receptor expression level significantly raised than that in 2 months (P＜0.05).Conclusions Long term ketamine addiction exerts toxicity not only on the bladder but also on the kidney.The increased expression of H1 receptor in rats' kidney and bladder tissues of group A indicates that H1 receptor may be related to the ketamine-associated urinary system dysfunction.The urine sodium and potassium within 24 h may be a sensitive index for the assessment of degree of kidney damage in the early stage of ketamine-induced dysfunction than serum creatinine.

Objective To investigate the correlation between ICRU reference point dose and dosevolume parameters of organs at risk (OARs) under different bladder and rectal filling status in threedimensional conformal brachytherapy for locally advanced cervical cancer.Methods A total of 31 patients who received magnetic resonance imaging-guided three-dimensional conformal brachytherapy for cervical cancer in 96 fractions were enrolled.The ICRU rectal and bladder reference points were determined in the treatment planning system,and the doses at these points were recorded and compared with the dose-volume parameters of the rectum and bladder.The paired t-test was used to analyze the differences between them.Results Bladder DICRU was lower than bladder D0.1cm3 and D1 cm3 (P=0.000 and 0.000),higher than bladder D5 cm3 and D10cm3 (P=0.000 and 0.000),and similar to bladder D2 cm3 (P=0.345).Under the bladder filling status,bladder DICRU was lower than D2cm3.Rectal DICRU was lower than rectal D0.1 cm3 and D1cm3 (P =0.000 and 0.002),higher than rectal D5 cm3 and D 10 cm3 (P =0.000 and 0.000),and similar to rectal D2cm3 (P=0.058).The ICRU bladder and rectal reference point doses were positively correlated with corresponding D2 cm3.In the case of bladder volume ≥ 200 cm3,the ICRU bladder reference point dose underestimated bladder D2 cm3.In the case of rectal volume ≥ 37 cm3,the ICRU rectal reference point dose overestimated rectal D2 cm3.Conclusions In three-dimensional conformal brachytherapy,it is generally safe to use D2 cm3 as an index to evaluate OARs,but when the bladder or rectum is in an empty status,the ICRU bladder or rectal reference point doses should be considered.

Objective To investigate the clinical results of the anterolateral thigh free flaps for the soft tissue coverage of Gustilo grade-Ⅲ B open bone fractures in lower extremities.Methods The anterolateral thigh free flaps were applied to treat 42 Gustilo grade-11Ⅲ B open fractures.Ten flaps were performed through an emergency procedure.Sixteen were performed at early stage and the other 16 were performed at later stage .The results of the therapy were analyzed after long time follow up. Results All limbs were salvaged and all the flaps survived without protracted course to obtain soft tissue coverage.The follow up ranged 11 to 47 months.The healing time of the bones were significant prolonged in the later stage therapy group compared with the emergency therapy group and the early stage therapy group.In the later stage therapy group,the incidences of the bone infection and nonunion were also higher than the other two groups,and the second stage bone transplantation to repair bone defect and nonunion were needed in 76% patients in this group.The lower limb functions of the later stage treated group were worse than those of the other two groups.Compared to the early stage treated group,better lower limb functions could be obtained in the emergency treated group. Conclusion Delayed soft tissue coverage resulted in higher incidence of complications.The immediate soft tissue coverage of severely injured limbs complicated by Gustilo grade-Ⅲ B open fractures and massive soft tissue defects had the advantages over traditional methods.Hard work though it was,one-stage soft tissue coverage using anterolateral thigh free flaps could obtain better lower limb function.