Immunogenicity of the truncated NDV F protein surface-displayed on Lactobacillus casei / 生物工程学报

To evaluate immune efficacy of the recombinant Lactobacillus casei, we constructed pLA-Newcastle disease virus (NDV)-F/L. casei and obtained the expression products. PCR amplified the NDV F gene carrying part of the major epitopes. The target gene was inserted to the shuttle plasmid pLA, and then transformed into Escherichia coli BL21 (DE3) in order to screen positive recombinant plasmid. The positive recombinant plasmid was transformed into L. casei by electroporation to construct pLA-NDV-F/L. casei. The positive strains were identified by PCR. The reactivity of the recombinant bacteria was identified by Western blotting and the protein expression was detected by indirect immunofluorescence, flow cytometry and laser confocal microscopy. The 14-day-old chickens in each group were vaccinated by oral plus nose drops. The pLA-NDV-F/L. casei twice immunization group and three times immunization group, the commercial vaccine group, the pLA/L. casei group, the unchallenge PBS and the challenge PBS group were established. IgG in serum and sIgA in the lavage fluid of intestinal, nasal and lung were detected by ELISA. The protection rate of chickens was evaluated. The results showed that 94.10% of the recombinant bacteria expressed the F protein. The recombinant protein was highly expressed on the surface of L. casei with a protein size of 62 kDa, which specifically bound to anti-NDV serum. The levels of anti-F IgG and sIgA antibodies in each test group were significantly higher than those in the control groups. The duration of antibody in the pLA-NDV-F/L. casei three-time immunization group lasted 28 days longer than that in the twice immunized group, and there was no significant difference between antibody peak values. The attack protection rates in each group of immunized pLA-NDV-F/L. casei three times, twice, attenuated vaccine, pLA/L. casei and PBS were 80%, 80%, 90%, 0% and 0%, respectively. Therefore, the antigenic protein of NDV F was successfully expressed by L. casei expression system, which has of reactogenicity and immunogenicity, and could induce protective immune responses in chickens.

Clinical features and treatment of residual gallbladder lesions after cholecystectomy / 中华消化外科杂志

Objective To investigate the clinical features and treatment of residual gallbladder lesions after cholecystectomy.Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 83 patients with residual gallbladder lesions after cholecystectomy who were admitted to the Affiliated Hospital of Inner Mongolia Medical University between January 2009 and April 2016 were collected.Among the 83 patients,74 had residual gallbladder (41 combined with bile duct stones and 33 with simple residual gallbladder)and 9 had simple residual bile duct stones.Patients received laboratory and imaging examinations,and then selected suitable surgery according to residual lesions.Observation indicators included:(1) clinical features:medical history,clinical manifestation,features of laboratory and imaging examinations;(2) surgical and postoperative situations:surgical procedure,operation time,volume of intraoperative blood loss,time of drainagetube removal,postoperative complications and duration of postoperative hospital stay;(3) postoperative pathological examination;(4) follow-up situation.Follow-up using outpatient examination and telephone interview was performed to detect survival of patients and recurrence of lesions using abdomen color Doppler ultrasound up to October 2016.Measurement data with normal distribution were represented as ~ ±s.Results (1) Clinical features:① medical history:all the 83 patients had histories of cholecystectomy,including 57 with emergency operation and 26 with selective operation.② Clinical manifestation:initial clinical manifestation occurred at 1-324 months postoperatively,with an average time of 96 months.Of 83 patients,49 had right upper abdominal colic associated with right shoulder or back rediating pain,fever,nausea and vomiting,24 had jaundice as the main symptoms,8 had gradually worse conditions,with right upper quadrant abdominal pain,chills and fever associated with jaundice in the advanced stage and 2 had abdominal discomfort associated with incomplete intestinal obstruction.③ Laboratory examination:elevated alanine transaminase (ALT) and aspartate transaminase (AST) levels were detected in 48 patients,elevated total bilirubin (TBil) level in 28 patients and elevated serum and urinary amylase level in 4 patients,respectively.④ Imaging examination:of 83 patients,77 had positive results of magnetic resonanced eholangio-pancreatography (MRCP),43 had positive results of abdominal color Doppler ultrasound and 39 (4 combined with pancreatitis) had positive results of abdominal computed tomography (CT).(2) Surgical and postoperative situations:all the 83 patients underwent surgery.Of 74 patients with residual gallbladder,72 underwent residual gallbladder resection (of 41 combined with bile duct stones,35 undergoing residual gallbladder resection + bill duct exploration and removing the stone + T-tube drainage,6 initially undergoing endoscopic sphincterotomy for removing bile duct stones and then residual gallbladder resection and 31 undergoing simple residual gallbladder resection),1 combined with residual gallbladder cancer underwent residual gallbladder resection + common bile duct resection + hepatic duct-jejunum internal drainage and 1 combined with residual gallbladder cancer and common bile duct space-occupying lesion underwent palliative biliary-enterostomy.Nine patients with simple residual bile duct stones underwent endoscopic sphincterotomy for removing bile duct stones.Operation time,volume of intraoperative blood loss,time of drainage-tube removal and duration of postoperative hospital stay were (92±39)minutes,(63±12)mL,(5±4)days and (9±5)days in 74 patients with residual gallbladder,respectively.There was no severe postoperative complication.Operation time,volume of intraoperative blood loss and duration of postoperative hospital stay were (57±33)minutes,(25±9)mL and (5 ± 3) days in 9 patients with simple residual bile duct stones,respectively.No severe complication was occurred in 9 patients.(3) Postoperative pathological examination:the length of residual cystic duct in 74 patients with residual gallbladder was (2.8 ± 1.0) cm,combining with stones.Of 74 patients,simple hyperplasia of residual gallbladder were detected in 54 patients,dysplasia in 14 patients,tumor-like hyperplasia and benign tumor in 4 patients and adenocarcinoma in 2 patients.(4) Follow-up situation:all the 83 patients were followed up for 6-71 months,with a median time of 33 months.Eighty-one patients were healthy survival,without recurrence of lesions.One patient had recurrence of bile duct stones at 34 months postoperatively,and then was cured by choledochojejunostomy,without recurrence up to the end of follow-up.One patient with residual gallbladder cancer complicated with common bile duct space-occupying lesion died at 9 months postoperatively.Conclusions Patients have recurrent right upper abdominal colic associated with right shoulder or back rediating pain,fever and jaundice after cholecystectomy,and lesions of residual gallbladder or bile duct stones should be considered.Reoperation is safe and feasible for patients,with a good clinical outcome.

Anti-tumor effects of p53N15-based fusion peptide in the transfected H1299 lung cancer cells / 中华胸心血管外科杂志

ObjectiveLoss-of-function mutation of p53,a tumor suppressor gene,is an important mechanism for the development of human cancers.In this study we tried to transfect p53N15-based fusion peptide into H1299,a lung cancer cell line,and evaluate the anti-tumor effects of the fusion peptide.MethodsAdeno-associated virus ( AAV) vectors were used for transfecting p53N15 fusion peptide into p53-null lung adenocarcinoma H1299 cells.The anti-replication effects of p53N15 fusion peptide were evaluated with inverted microscopy,MTT test for cell viability and flow cytometry.ResultsFusion peptides in H1299 cells was highly expressed and had detectable suppressive effects on cell proliferation.A large amount of dead cells were seen under microscope after the transfection of recombinant viruses for 72 hours.Cells activity was reduced significantly in the virus-transfecting groups as demonstrated by MTT test.The flow cytometry showed that a large number of dead cells were present in the virus-transfecting groups.ConclusionThe growth of H1299 lung adenocarcinoma cells could be inhibited in vitro by being transfected with p53N15 fusion peptide,which may be a potential gene therapy alone or as an adjuvant option in the treatment of lung cancer.