Objective To obtain sufficient recombinant multiepitope peptide of ESAT-6.Methods DNA encoding ESAT-6 peptide was amplified by PCR using a pair of primers which were designed in accordance with the reported ESAT-6 gene sequence.The ESAT-6P gene was inserted into the prokaryotic expression vector pET-30a with the N-terminal 6His-tag.The recombinant peptide vector pET30a-ESAT-6P was transformed into E.coli BL21(DE3) via chemical transformation.The positive clone was screened by means of PCR.The target peptide was expressed in E.coli after induction with IPTG and analyzed by SDS-PAGE.The immunogenicity of the peptide was evaluated by dot immunobinding assay.Results The target peptide was successfully expressed and purified.The solubility analysis showed that the recombinant peptide existed as inclusion body in the E.coli.DIBA indicated that the ESAT-6P was specifically reactive to sera from TB patients.Conclusions The recombinant multiepitope peptide of ESAT-6 engineering bacteria has been obtained,which will be quite helpful to develop novel specific diagnostic reagents and effective vaccine.

Objective To evaluate the clinical value of laparoscopy in the diagnosis and treatment of intestinal duplication for children. Methods From June 2002 to May 2007, 15 children (aged 27 days to 10 years) with intestinal duplication were treated by laparoscopy in our hospital. The lesions were explored by laparoscopy, and then pulled out through the umbilical hole or the trocar hole at the right lower abdomen. The anastomosis was performed outside the abdominal cavity. Results The operation was successfully accomplished in all the patients. No serious postoperative complications occurred. Among the cases, end-to-end anastomosis was performed on 12 patients, resection of the duplicated intestine on 2, and demucosation on 1. Simultaneously, appendectomy was performed on 2 cases, and ring suturing for inguinal hernia on 1. The patients were followed up for 3-18 months (mean, 12 months), during which no patients had symptoms of intestinal adhesion (abdominal distension, vomiting), hemorrhage, or incisional hernia. Conclusions Laparoscopy is effective and reliable with a high diagnostic rate and low complication rate in the diagnosis and treatment of intestinal duplication for children.

OBJECTIVETo investigate the clinical application of expanded flap based on cutaneous branch of transverse cervical artery for reconstruction of cervical cicatricial contracture.

METHODSBased on the clinical anatomy of cutaneous branch of transverse cervical artery flap, we design the corresponding subclavicular area for expansion. The incision was usually located at the anterior axillary fold, 5-8 cm in length. The expander was implanted under the deep fascial layer, without injury of the vascular pedicle. Fixation sutures were put about 1 cm apart from the incision to prevent the expander from transposition and exposure. After expansion, the cervical cicatricial contracture was excised and released. According to the defect, the expanded flap based on the cutaneous branch of transverse cervical artery was designed, with the pedicle located at the posterior margin of sternocleidomastoid and 1. 8 cm above median point of clavicle. "S" shape incision was made at the location of vascular pedicle. Subcutaneous dissection was performed 1.5 cm in width along the incision on both sides. Then the flap was harvested under the deep fascial layer and rotated to cover defect without tension. It was not necessary to dissect the vascular pedicle further. The defect at donor site was closed directly.

RESULTS17 cases were treated with the island flap. The contracture of the cervical scar was corrected completely with aesthetic appearance.

CONCLUSIONSExpanded flap based on cutaneous branch of transverse cervical artery has reliable blood supply. It' s an ideal flap for the treatment of cervical cicatricial contracture.

Arteries ; Cicatrix ; surgery ; Clavicle ; Contracture ; surgery ; Dissection ; methods ; Fasciotomy ; Humans ; Neck ; blood supply ; Neck Muscles ; anatomy & histology ; Surgical Flaps ; blood supply

Objective To explore the method of total nasal reconstruction when the forehead skin for expanding is unavailable.Methods According to the principle of total nasal reconstruction,total nasal scar and deformity were repaired with expanded random forearm falp.All the expanders were placed in flexor side of forearm.The incision sides were placed in proximal,distal,or lateral part of the forearm according to different pedicles.After expansion,the nares were enlarged,eversion of ala nasi corrected,the contracture scars sufficiently released,and the size and shape of the reconstructive nose designed according to face size.The scar of nasal dorsum and capsule of the expanded flap could be used for reconstructing nasal dorsum.Donor sides could be sutured directly,the pedicle could be cut 3 weeks later.Results All the flaps survived with good appearances,and 3 of the 15 cases with proximal,11 with distal,and 1 with lateral pedicle.The effect of distal pedicle group was better than that in proximal and lateral pedicle group because of comfortable posture longer pedicle,and providing more tissue for reconstruction.Conclusions Total nasal reconstruction with expanded random forearm flap is an option when the forehead skin for expanding is unavailable.

For a matured digestive surgeon,pancreaticoduodenectomy (PD) is regarded as one of the most complicated and technically challenging surgical procedure.Based on the accurate interpretation of patient's preoperative imageologic data,we advocate a novel procedure which is called as G-path pylorus-preserving pancreatoduodenectomy (G-path PPPD).We deen G-path PPPD as a standardized procedure for resectable pancreatic head cancer or periampullary carcinoma,which definitely simplify the procedure,save the operative time,achieve R0 resection through en-bloc resection without interruptedly intraoperative exploration and reduce the risk of iatrogenic tumor metastasis.This article introduced the program of G-path PPPD in detail by taking a patient as an example who suffered from pancreatic head cancer accompanied with obstructive jaundice,and discussed the relevant points.

Objective To investigate the treatment methods of tissue defects in nose and lower eyelid area.Methods Based on the impaired area,combined pre-expansion of forehead and facial region was adopted.The volume of soft tissue expander was determined.150-170 ml expander was implanted between frontalis muscle and periosteum of forehead,while at the facial region,and the volume of expander should be determined by the normal skin area as large as possible.The excision of impaired tissue was performed after sufficient expansion,and then skin flap or transposition skin flap were advanced at the facial region.At forehead,ortho-position skin flap based on supratrochlear vessels was designed for repairing the nose and palpebra inferior region.Three weeks later division of the ortho-position skin flap was carried out.With the flattened pedicle,the rest part of impaired area was replaced.Results Six cases were performed with this method.For all the cases,the survival rate of flap was 100％.Esthetic appearance and satisfying color-match were achieved,without appearance of lower eyelid ectropion.Conclusions Combined pre-expansion of forehead and facial region is an ideal method in treatment of the defects in nose and lower eyelid area.

Objective To investigate the feasibility and outcome of resection and reconstruction of hepatic artery in hilar cholangiocacinoma (HCC).Methods The data of 29 patients with HCC with hepatic artery reconstruction carried out from March 2009 to August 2013 in our center were retrospectively analyzed.23 right hepatic arteries and 6 common hepatic arteries were involved.In-situ anastomosis was carried out in 20 patients and a double anastomosis using gastrodoudenal artery grafts was carried out in 9 patients.Results There were no arterial thrombosis or other related complications on prolonged follow-up.Conclusion Hepatic artery resection and reconstruction should be carried out if the artery was invaded by a hilar cholangiocarcinoma to produce a high resection rate and a better outcome.

Objective To investigate the expression of programmed cell death-1(PD-1)ligands on HepG2 cell and 2.2.15 cell.Methods Culture HepG2 cell and 2.2.15 cell in vitro and stain the cells with fluorescence-labeled monoclonal antibodies specifically against PD-L1 and PD-L2,respectively.Isolate total RNA of HepG2 cell and 2.2.15 cell and detect the mRNA expressions of PD-L1 and PD-L2 by RT-PCR.Results The expression of PD-L1 on 2.2.15 cell was upregulated on the levels of both proteins and mRNA.The expression of PD-L2 in 2.2.15 cell was not detected.And neither the PD-L1 nor PD-L2 was detected in HepG2 cell.Conclusion HBV infection can upregulate the expression of PD-L1 on hepatocyte,which might contribute to the immune evasion of HBV.

Objective To explore the clinical features of methylmalonic acidemia (MMA) in children admitted to the pediatric intensive care unit, to help improve our understanding of MMA. Methods The clinical data of 21 patients with MMA admitted to our PICU from December 2012 to August 2016 were analyzed. Diagnosis were confirmed by gas chromatography-mass spectrometry, GC/MS. Results twenty-four of 158 suspected cases were confirmed as having organic acidemia diseases including 21 cases of MMA, one case of propionic acidemia, one case of urea cycle disorders, and one case of glutaric acidemia. The main clinical manifestations were feeding difficulty, malnutrition (13 cases), developmental retardation (12 cases), lethargy (10 cases), tricuspid severe reflux and pulmonary hypertension (1 case), hydrocephaly (5 cases), muscular dystonia (three cases with hypertonia, and four with hypotonia), convulsion (7 cases), apnea, sobbing respiration (10 cases), chromatosis (6 cases), anemia (13 cases), edema (6 cases), thrombocytopenia (6 cases), hematuria and proteinuria (2 cases). Five cases gave up therapy before diagnosis was made. Sixteen cases received the treatment with Vitamin B12 and supplementation of L-carnitine. Seven cases gave up after treatment without effect or deterioration of condition. Eight cases were vitamin B12-responsive, and one case was vitamin B12-nonresponsive. The follow-up for a period ranging from three months to two years, among eight vitamin B12-responsive cases, 6 cases showed a favorable outcome with apparent improvement, one case had no symptom and one patient died from severe pneumonia. Vitamin B12-nonresponsive case was still alive. Conclusions The clinical manifestations of MMA are non-specific. Urine organic acid analysis is critical to early diagnosis of MMA in high-risk patients. Timely diagnosis and appropriate long-term treatment are essential to improve the prognosis of the disease.

Objective To summarize the initial experience in adult-to-adult living donor liver transplantation(ALDLT),so as to improve the efficacy of ALDLT.Methods The clinical data of 31 adult patients who undelwent ALDLT from June 2006 to February 2008 were retrospectively analyzed.Results Of all the patients,8 was with decompensated cirrhosis,7 with acute liver failure,12 with hepatocellular carcinoma,2 with purpura of liver,1 with hilar cholangiocarcinoma,and 1 with Wilson disease.The liver grafts included 25 right lobes with middle hepatic vein(MHV),3 right lobes without MHV,1 right lobe with MHV+left lateral lobe,1 right lobe with MHV+left lobe.The remaining 1 patient underwent auxiliary partial orthotopic liver transplantation with left lobe graft.Six post-transplantation complications occurred in 5 donors. Eleven post-transplantation complications occurred in 9 recipients,including 4 biliary complications,3 vascular complications,3 infection complications and 1 delayed healing of the incision.After ALDLT,2 recipients died of pulmonary infection and 1 of multiple aspergillus infection. Conclusions ALDLT has become an effective method to expand the source of liver grafts.Rational donor and recipient assessment,surgical procedure and postoperative management are key to ALDLT.