Objective:To clone and express active domain of human granzyme A ( aGzmA ) and detect its biological activity.Methods:Human aGzmA gene was amplified by PCR from the full-length human granzyme A and inserted into prokaryotic ex-pression vector pET24a(+).The constructed recombinant plasmid pET24a-aGzmA was transformed to E.coli BL21(DE3) and induced with IPTG.The expressed product was identified by SDS-PAGE and Western blot.The recombinant protein was purified by the Ni2+affinity column chromatography and the enzyme activity was assayed with BLT substrate.Results: A DNA fragment of 700 bp was amplified by PCR.The recombinant plasmid pET24a-aGzmA identified by enzyme-digesting analysis and sequencing showed that aGzmA gene was inserted into vector plasmid correctly.SDS-PAGE analysis showed that there was a specific protein with a relative molecular mass of about 26 kD.Western blot analysis indicated that the protein could react with mouse anti-His monoclonal antibody specifically.The recombinant protein with high purity could be acquired from the inclusion bodies by the Ni2+ affinity column chromatography and the purified protein had good enzyme activity.Conclusion: The recombinant human granzyme A with good biological activity was prepared successfully.

Clinically, kidney?yang deficiency is one of the main TCM syndromes in patients with major depressive disorder. Warming yang and reinforcing kidney as a therapeutic measure for the treatment of depressive disorder has a?chieved good clinical curative effect. However, there are some problems on the study of kidney?yang deficiency depression. One of the reasons is that the establishment methods of depression animal model with kidney?yang deficiency are not uni?fied. In order to provide reference and further improve the replication method of depression animal models with kidney?yang deficiency, we systematically collected and analyzed the experiment results published in the past 15 years about the depres?sion animal model with kidney?yang deficiency induced by glucocorticoid hormone drugs and find out their similarities and differences.

The article presents a method improvement of the medial plantar pedicled flap transplantation to cover the defect of the foot heel.Since 2002,6 patients with soft tissue defects on the foot heel underwent reconstruction with the medial plantar pedicled flap transplantation of excluding the plantar fascia and 3 patients used conventional flap of the medial plantar pedicled flap transplantation.They ranged in age from 18 to 52 years(mean,32 years).All the flaps had survived completely without major complication with satisfactory clinical results.Follow-up period ranged from 1.5 to 6 years(mean,3.8years)postoperatively.There was no remarkable donor site morbidity.Patients with flap of excluding the plantar fascia had good appearance in recipient site.Injury to donor site was reduced.Skin of the instep area can be raised as an island fasciocutaneous flap based on medial plantar vessels,with the branch of medial plantar nerve supplying the instep skin to provide the sensation.The flap is thin and suitable for repairment of soft tissue defect on the foot heel.

ObjectiveTo evaluate the clinical efficacy and safety of nylon ligation combined with endoscopic high-frequency electric snare for the treatment of colorectal polyps.MethodsThe clinical data of 345 patients with colorectal polyps who received the treatment of nylon ligation combined with endoscopic high-frequency electric snare at the Zhongshan Hospital of Fudan University from January 2006 to January 2011 were retrospectively analyzed.Complications including bleeding and perforation were observed.Postoperative wound healing and local residual recurrence were assessed during follow-up.ResultsA total of 362 colorectal polyps were successfully removed.The mean diameter of the colorectal polyps was 2.7 cm (range,1.5-4.0 cm). Operative bleeding occurred in 1 patient,and perforation in 1 patient.The results of histological examination showed that 93.6％(339/362) colorectal polyps were adenoma,17 were hyperplastic polyps,5 were inflammatory polyps,and caneeration of colorectal polyps in 1 patient was detected.Endoscopic follow-up was completed in 334 patients at the first month after operation,and 9 patients with residual benign colorectal polyps received endoscopic resection.No residue or recurrence was found in the other patients.Endoscopic follow-up was completed in 308 patients at the second months after operation,and no residue or recurrence was found.The overall effective rate was 97.1％ (299/308).ConclusionEndoscopic nylon ligation combined with endoscopic high-frequency electric snare is effective and safe for the treatment of colorectal polyps.

Objective To evaluate the clinical efficacy and safety of metallic hemoclips in the treatment of gastric defects during endoscopic full-thickness resection. Methods Patients with submucosal tumors derived from the muscularis propria and adhesion of the gastric serosa diagnosed by EUS and CT were enrolled in the study.A total of 62 patients,22 males and 40 females,mean age 58.5 years,were recruited into the present study from June 2009 to December 2010,in which 37 patients were with tumors in gastric fundus,20 in gastric body and 5 in antrum.All patients were treated with endoscopic full-thickness resection.After the operation,metallic hemoclips were used to close the defects through endoscopic biopsy channel.The closure success rate with metallic hemoclips were assessed.The patients were followed up endoscopically for evaluation of efficacy and safety of the procedures.Results All patients underwent endoscopic full-thickness resection.Uncontrollable bleeding occurred in 1 patient,who subsequently underwent emergent laparoscopic surgery and was excluded from the study.Metallic hemoclips were applied to close defects in 57 patients,and metallic hemoclips combined with omentum obstruction in 4 patients.The overall success rate was 100％ in 61 patients.Endoscopic follow-up was completed in all patients in average of 4.4 months.The clinical wound healing rate was 100％.No complications such as fever,abdominal pain were found in 2months after the procedures.Conclusion Application of metallic hemoclips during endoscopic full-thickness resection for gastric defects is a safe and effective technique.

Objective To summarize clinical experience in application of nail-bed lengthening operation for fingertip injuries.Methods From September 2003 to May 2007,36 patients (25 men and 11 women) aged 16 -45 years (26 in average) with nail defect by fingertip injuries underwent reconstruction operation using nail-bed lengthening at Orthopaedics and Traumatology Hospital of Taizhou,with the thumb involved in 15 cases,the index finger in 10 cases,the middle finger in eight eases and the ring finger in three case.A skin rectangle with thickness of 2 - 3 mm and width as that of the nail was resected at the skin edge 5 -6 mm distant from the epenychium,without damage to underlying subcutaneous vascular network, and U-shape skin flap was delicately slid proximally with its ends sutured.Results All surgical operations for nail lengthening were uneventful and clinically satisfactory,and the appearance of the thumb or other fingers was good during follow-up for six months to three years (19 months in average).Conclusions Nail lengthening for fingertip injuries with nail defect is a simple and effective surgical operation which could improve the appearance of the thumb and other fingers.

The present article reports the method of preparation of epidermal growth factor (EGF) from mouse submaxillary glands by using a two-step liquid chromatography and the efficacy of EGF in promoting animal wound healing. It was found that combined purification of crude EGF is better with Bio-gel p-10 and DEAE-DE52 ion-exchange chromatographies and purified EGF has a marked enhancement of wound healing and reduction of cicatricial contracture in animals.

Aim To establish depression rat model with kidney-yang deficiency and evaluate the related parameters.Methods A total of 24 adult SD rats were divided into control group,mod-el group and self-healing group by body weight.Model group and self-healing group were injected hydrocortisone for 21 days, meanwhile the control group was injected sodium chloride.After modeling,the related indexs detection were taken out:open-field test,sucrose preference test and forced swim test.Then the orbital blood was taken to detect 5-HT,DA and NE by LC-MS/MS.And liver,kidney,spleen,thymus,adrenal gland,testis and epididymis were taken to calculate organ index.Results Compared with the control group,model rats displayed depres-sion and kidney yang deficiency syndrome such as dry hairs, chills,back arched and so on. Correspondingly, the body weight increased slowly.Immobility in FST was significantly in-creased. Sucrose preference, horizontal scores and vertical scores was significantly decreased.Organ index decreased in liv-er,kidney,spleen,adrenal,testis and epididymis.The level of 5-HT and NE was significantly high.Compared with the model group,5-HT,NE and DA were significantly reduced,5-HT and DA were even lower than those of the control group.Conclusion Depression rat model with kidney-yang deficiency could be in-duced by continuous injection of hydrocortisone.

Objective To summarize clinical experience of application of muliplefoliated flap combined with tendon graft from dorsal foot for repairing soft tissue defects of multi-fingers. Methods Based on the results of microdissection researches, muliplefoliated flap including medial foot flap, the first metatarsal dorsal flap, lateral foot flap, extensor pollicis brevis of toe and extensor proprius of the fifth toe pedicled by anterior tibial vessels and dorsal pedis vessels as trunk, anterior medial malleolus vessels, the first metatarsal vessels and anterior lateral malleolus vessels as branches respective was designed for repairing soft tissue and tendon defects of multi-fingers. Results In all 12 cases, total survival of flap was 11 cases , 1 case had partial necrosis of distal flap and it was healed by dress changing. Conclusion Transplantation of muliplefoliated flap combined with tendon graft from dorsal foot is a good method to repair soft tissue and tendon defects of multi-fingers.

Objective To investigate the clinicopathological features of specific subtypes of laterally spreading tumor (LST) and assess the outcomes of endoscopic submucosal dissection (ESD) based upon differentiation status.Methods A total of 174 cases of colorectal LST,treated with ESD between January 2007 and April 2011,were divided into granular type (LST-G) and non-granular type ( LST-NG),and further divided into 4 subtypes,including homogeneous granular,nodular mixed,flat elevated and pseudo-depressed types.Clinicopathological data and ESD parameters were compared between the four subtypes.Results Nodular mixed tumors were larger than homogeneous tumors,needed longer operation time and were more frequently seen in malignant transformation,high grade intraepithelial neoplasm ( HGIN ) as well as submucosal invasive cancer.So were peeudo-depressed tumors compared with flat elevated ones ( P ＜0.01 ).There was no significant difference between LST-G and LST-NG in the parameters of ESD including en bloc resection rate,complete resection rate and complete cure resection rate.The complications tended to be more frequent in nodular mixed tumors (bleeding rate:4.6％; peroration rate:3.5％ ) and pseudo-depressed tumors ( bleeding rate:11.1％ ; perforation rate 11.1％ ),but there was no severe complication in either homogeneous tumors or fiat elevated tumors.One recurrent pseudo-depressed tumor was found 6 months after ESD treatment.Conclusion The degree of technical difficulty appears higher for nodular mixed and pseudo-depressed tumors than for homogeneous and flat elevated tumors,as is shown in procedure time and complication rate.ESD for these two subtypes should better be performed by experienced endoscopists.Accurate differentiation of the LST lesions is needed in basic and clinical research.