Objective Construct an expression vector for anti-HBs Fab fragment and IFN-? fusion protein, then transform it into E.coli Top10 in order to produce a targeting drug for hepatitis B. Methods Using PCR and molecular clone techniques, we amplify the gene fragment of IFN-? with corresponding endonuclease sites and artificial linker at 5′,3′ termini,then recombine it within the vector of anti-HBs Fab gene in correct endonuclease sites at 3′ termini of the Lc gene, choosing the positive clone with PCR and transform it into E.coli Top10,after the expression and purification of the fusion protein, we use ELISA and Dot-blot to verify the antigenicity and binding activity to HBsAg. Results We got fusion protein consisting Lc and IFN-?. It has the IFN-? activity and HBsAg affinity which is similar to anti-HBs Fab fragment. Conclusions The successful expression of the fusion protein with both HBsAg affinity and IFN-? activity make it possible for immunobiotherapy drug targeted to HBV.

The purpose of this study was to compare the efficiency of 3 different methods in purifying engineered anti-HBsFab. Anti-HBsFab Supernatants containing anti-HBsFab were prepared by ultrasound lysis of host cells of anti-HBsFab positive clone. Anti-Fab-sepharose gel, streptococcal protein G-sepharose gel (Prot G gel) and Ni-NTA-agarose gel were used to purify Fabs according to the reagents protocols. Then the concentrations of purified proteins were determined. SDS-PAGE was used for the measurement of purified Fabs'purity. HBsAg based ELISA was chosen to determine the Fabs' bio-activity. The results indicated that Fab recovery of different gels in equal volume were different. The recovery of Ni-NTA GEL was greater than that of Prot G gel, and the recovery of Prot G gel was greater than that of anti-Fab gel. Purity of the Fab isolated by 3 different gels was confirmed by SDS-PAGE. The binding capacity of anti-HBs Fab to HBsAg of these three gels had no signifticant difference. Our data suggested that each method had its advantage and usage limitation, Ni-NTA method demonstrated much better performance in economy, management, and efficiency over the other two methods.

Objective:To show whether the Fas Ligand gene induces mast cells apoptosis.Methods:RT-PCR was used to amplify the gene of rat Fas ligand extracelluar domain and transmembrane domain and cloned it into eukaryotic expression plasmid pcDNA3.1.Transcent transfect RBL-2H3,the expression of Fas ligand RBL-2H3 was detected by RT-PCR、Western blot.The Annexin V FCM was used to detect the RBL-2H3 apoptosis after the transfection of Fas Ligand.Results:It is successful to obtain the gene of rat Fas Ligand extracellular domain and transmembrane segment,cloning it into pcDNA3.1,FasL was expressed on the surface of RBL-2H3 and it's supernatant after the transfection of pcDNA3.1/FasL.The cell start to be apoptosis.Conclusion:Our study reveals that Fas Ligand gene transfection in RBL-2H3 can effectively induced apoptosis.It is a promising strategy for Fas Ligand to be used in the therapy of allergic disease. [

OBJECTIVETo improve the treatment for severe open tibia-fibular fracture.

METHODSFrom 1994 to 2000, 146 patients with severe open tibia-fibular fracture were treated. According to Gustilo classification, all patients were of type III. Among them, 96 patients belonged to III A, 36 III B, and 18 III C. One hundred and eight patients were male and 38 female, aged from 11 to 68 years, with an average of 31. All patients were treated with unilateral external fixator combined with simple internal fixation (general screw or Kirschner wire). Thirty patients were treated with secondary flap operation. Among them, 19 patients received pedicle gastrocnemius muscle flaps, 9 free vastus lateralis muscle flaps, and 2 free latissimus dorsi muscle flaps.

RESULTSThree patients of type IIIB were subjected to amputation because of advanced age and associated cerebral or thoraco-abdominal injury. Five patients of type III C had amputation because of insufficient postoperative blood supply and necrosis. The rupture of other 138 patients was well reduced, and firmly fixed. They were followed up for 6 months-6 years, with an average of 2.5 years. The average time of fracture-union was 27 weeks, and the average time for removal of fixtors was 28 weeks. The motion of knee joint ranged from 0 to 120 degree in 110 patients; from 0 to 100 degrees in 25, and from 0 to 90 degrees. The motion of ankle joint was approximately normal.

CONCLUSIONSFor patients with severe open tibia-fibular fracture, comprehensive analysis should be made for preservation of the wounded limb or amputation as for elderly patients with vessel-nerve injury or with cerebral- thoracoabdominal injury, emergency amputation should be done. Unilateral external fixator combined with simple internal fixation (general screw or Kirschner wire) for severe open tibia-fibular fracture is advantageous for a simple and reliable fixation. It is less traumatic.

Adolescent ; Adult ; Aged ; Child ; External Fixators ; Female ; Fibula ; Fracture Fixation, Internal ; Fracture Healing ; Fractures, Open ; surgery ; Humans ; Male ; Middle Aged ; Tibial Fractures ; surgery

BACKGROUND: With the rapid spread of novel coronavirus pneumonia (NCP) worldwide and the escalation of prevention and control efforts, the routine medical needs of patients have been restricted. The aims were to investigate medical needs of lung cancer patients and their mental health status during the epidemic periods, so as to provide rational recommendations for subsequent diagnosis and treatment. METHODS: The questionnaire was sent in the form of an electronic questionnaire at 7am on 4th, March, 2020, until 7am 6th, March, 2020, 368 questionnaires were recollected from 25 provinces (autonomous regions/municipalities) in 48 h. RESULTS: Of the 368 patients, 18 patients were excluded as they didn't receive anti-tumor treatment, and 350 patients were included in the final analysis. 229 cases were treated with oral targeted drugs, and 121 cases were treated with chemotherapy or immunotherapy. 41.3% of patients treated with intravenous chemotherapy or immunotherapy experienced treatment discontinuation, and the proportion of treatment discontinuation in chemotherapy or immunotherapy was higher than those treated with oral targeted drugs (21.0%). Whether oral targeted drugs or intravenous chemotherapy or immunotherapy, more than 60% of patients experienced delays in imaging examinations. Nearly one third of patients developed new symptoms or exacerbation of existing symptoms. 26.6%-28.9% of patients have changed their treatment plans through online consultation. During novel coronavirus pneumonia, 40%-75% of lung cancer patients have mental health problems, and more than 95% of patients support government's prevention and control measures. CONCLUSIONS During the emergence of NCP, the medical needs of patients with lung cancer have not been enough, especially those who discontinued chemotherapy or immunotherapy. When medical institution resumes work, priority should be given to them. At the same time, mental health problems of patients should be valued and resolved timely.
Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Betacoronavirus ; physiology ; China ; epidemiology ; Coronavirus Infections ; epidemiology ; virology ; Female ; Humans ; Lung Neoplasms ; drug therapy ; psychology ; Male ; Middle Aged ; Pandemics ; Pneumonia ; epidemiology ; virology ; Pneumonia, Viral ; epidemiology ; virology ; Retrospective Studies