To enhance the specificity of anti-TNF-α single chain Fv antibody (TNF-scFv) to inflamed site, we constructed a bispecific antibody BsDb that targets TNF-α and ED-B-containing fibronectin (B-FN) by covalently linking TNF-scFv and the anti-ED-B scFv L19 at the gene level via a flexible peptide linker deriving from human serum albumin. BsDb was successfully secreted from Pichia pastoris as functional protein, identified by immunoblotting, and purified to homogeneity with affinity chromatography. BsDb retained the immunoreactivity of its original antibodies TNF-scFv and L19, and showed a marked gain in antigen-binding affinity and in TNF-α-neutralizing ability, when compared to TNF-scFv and L19 that were produced in Escherichia coli. In the adjuvant-induced arthritis (AIA) mice model, BsDb showed selective accumulation and retention in the inflamed paws but rapid clearance from blood, resulting in high arthritic paw to blood ratios. These data indicate that BsDb is endowed with high specificity to inflamed site and low toxicity to normal tissues and holds great potential for in vivo application for the targeted therapy of RA and other chronic inflammatory diseases.
Animals ; Antibodies, Bispecific ; biosynthesis ; immunology ; Antibodies, Neutralizing ; biosynthesis ; immunology ; Escherichia coli ; Fibronectins ; chemistry ; immunology ; Humans ; Mice ; Single-Chain Antibodies ; biosynthesis ; immunology ; Tumor Necrosis Factor-alpha ; immunology

In order to enhance the specificity of TNF-α monoclonal antibody to inflamed site, a bispecific antibody BsDb that targets TNF-α and the extra-domain B (ED-B) of fibronectin (FN) was constructed by covalently linking the anti-TNF-α single chain Fv antibody (TNF-scFv) and the anti-ED-B scFv L19 via a flexible peptide linker deriving from human serum albumin (HSA). ED-B is an antigen specifically expressed at the inflamed site. BsDb is expressed in E. coli, identified by immunoblot, and purified with affinity chromatography. This was followed by further examination of its bioactivities and pharmacokinetics. We demonstrated that BsDb retained the immunoreactivity of its original antibodies as it could simultaneously bind to TNF-α and ED-B and neutralize the biological action of TNF-α. In the collagen-induced arthritis mice model, BsDb selectively accumulate in the inflamed joint with a maximal uptake of (12.2 ± 1.50)% ID/g in a single inflamed paw and retain in the inflamed paw for at least 72 h. In contrast, BsDb showed a short serum half-life of (0.50 ± 0.05) h and a rapid clearance from normal tissues. The findings reported herein indicate that BsDb has good specificity to the inflamed site and low toxicity to normal tissues. BsDb is therefore likely to have greater clinical applications in the treatment of rheumatoid arthritis and other autoimmune diseases. This laid a stable basis for its preclinical study.

In order to investigate the species and categorization of Gasterophilus in Ili horse.We analysised the COI gene of the identified Gasterophilus dominant species and constructed NJ phylogenetic tree in the study.The results showed that infection rate was 100％ in total of 16 775 the third phase Gasterophilus instar larvae.Four Gasterophilus species were identified,and showed serious mix infections.Dominant species were Gasterophilus nasalis,its relative dominance were 53.17％,and prefer to live in the cardia,others to irregular live in the pylorus of the horses.COI gene homology of GasterophiIus nasalis,Gasterophilus intestinalis,Gasterophilus pecorum,Gasterophilus haemorrhoidalis (GenBank Accession No.:GU265752.1,KR230402.1,KU578262.1,KT946620.1) were 99％,99％,99％ and 100％ respectively.Phylogenetic analysis results showed that the data were clustered with the Gasterophilus app.which publshed on the GenBank.G.intestinalis and G.haemorrhoidalis cluster together first,and then cluster with G.nasalis,at last all three kinds of Gasterophilus cluster with G.pecorum.When the COI gene is the target,in-group and out-group of the Gasterophilus can forms an independent evolutionary branch.This study provides useful parameters for the classification of Gasterophilus.

Objective: This study aimed to explore the relationship between childhood sexual abuse (CSA) and sexual orientation among college students, and to explore possible sex difference. Methods: By using multi-stage stratified cluster sampling method, 4 034 students were selected from 4 college schools. Self-made questionnaire was used to collect the demographic information, CSA experiences and sexual orientation. Logistic regression models were conducted to examine sex differences in the relationship between different types and timing of CSA and sexual orientation. Results: The reporting rates of heterosexual, homosexual, bisexual and asexual orientation of college students were 93.2%, 0.7%, 3.7% and 2.4%, respectively. For males, contact CSA (OR=14.70, 95%CI=5.73-37.72), both contact and noncontact CSA (OR=4.33,95%CI=1.91-9.84) in elementary school or earlier were associated with sexual orientaion. non-contact CSA (OR=4.20, 95%CI=2.21-7.98), both contact and noncontact CSA (OR=3.57, 95%CI=1.65-7.70) in middle school were related to sexual orientation. However, for females, non-contact CSA (OR=1.78, 95%CI=1.02-3.13) and both contact and non-contact CSA (OR=3.13, 95%CI=1.35-7.23) in elementary school or earlier were associated with sexual orientation. Conclusion CSA experiences are associated with sexual orientation in sex-specific manner, with significant stronger association among males.

Chronic myeloid leukemia (CML) is a malignant tumor formed by clonal proliferation of bone marrow hematopoietic stem cells. With the improvement of disease awareness and the introduction of new drugs, more than 90% of CML patients can achieve long-term survival. However, a few patients still show drug resistance. This article reviews the mechanism of drug resistance in CML patients treated with tyrosine kinase inhibitor (TKI) and the characteristics of ABL kinase region mutation.

Objective:To investigate the clinical characteristics and prognosis of acute myeloid leukemia (AML) patients with positive TLS-ERG fusion gene.Methods:The clinical data of 9 AML patients with positive TLS-ERG fusion gene in the First Hospital of Jilin University from June 2013 to August 2020 were retrospectively analyzed, and the relevant literature was reviewed.Results:Among 9 patients with positive TLS-ERG fusion gene, there were 5 males and 4 females, with a median age of 16 years old (6-40 years old). Five patients received chemotherapy alone, 3 patients received allogeneic hematopoietic stem cell transplantation (allo-HSCT), and 1 patient did not receive systematic treatment. Among 8 patients with systematic treatment, 1 patient had complete remission after the first induction chemotherapy and 5 patients had complete remission after induction therapy. The median overall survival time of 5 patients with chemotherapy alone was 1.5 months (1-11 months), of which 3 patients did not respond to the first course of treatment and died of infection, and 2 patients died after relapse. The median overall survival time of 3 patients with allo-HSCT was 16 months (13-17 months), of which 2 patients died after relapse and 1 patient had sustained molecular complete remission by the end of follow-up.Conclusions:AML with positive TLS-ERG fusion gene has low incidence rate and poor induction efficacy. Hematopoietic stem cell transplantation may partially improve the survival prognosis of patients, but it cannot overcome the adverse effect of positive TLS-ERG fusion gene on prognosis.

Objective To analyze the exposed dose of hippocampus(HC)of T3,T4nasopharyngeal carcinoma patients treated with intensity modulated radiotherapy(IMRT). Methods The bilateral HCs were delineated and were divided into head(HH),body(HB)and tail(HT)for 62 nasopharyngeal carcinoma patients treated with IMRT.The dose parameters of HC were then analyzed. Results The mean dose of left and right HC was(1 127±704)cGy,(1 173±762)cGy. The mean dose of left HH,HB and HT was(1 732±1029)cGy,(820±632)cGy,(423±366)cGy(P=0.000);while the mean dose of right HH, HB and HT was(1 985±1101)cGy,(837±531)cGy,(432±343)cGy(P=0.000).The exposed dose and the volume exposed in different dose of HH were obviously higher than those of HB and HT.The dose parameters of HH,HB and HT decreased in turn. The involvement of sphenoid sinus,ethmoid sinus and cavernous sinus correlated with high exposed dose of HC. Conclusions The exposed dose of HH,HB and HT was different in nasopharyngeal carcinoma patients treated with IMRT.The exposed dose of HH was the highest,which should be emphasized especially. The involvement of sphenoid sinus,ethmoid sinus and cavernous sinus suggest high exposed dose of HC.

Inflammatory cytokines can mediate many biological processes and are tightly regulated by the body. Loss of control can trigger a range of diseases such as autoimmune inflammation and cancer. Therefore, a number of biological agents that can effectively regulate the biological effects of inflammatory cytokines such as recombinant anti-inflammatory cytokines, cytokine receptors and neutralizing antibodies have been extensively used in the treatment of related diseases caused by the imbalance of inflammatory cytokines. In recent years, in particular, a number of new innovative biological agents for blocking and regulating cytokine activities are emerging. In this article, we review the recent development and clinical use of the biologics targeting TNF-α, IL-1, IL-6 and IL-17, and point out their inherent limitations and clinical risks. Finally, based on the research findings of our own and other scholars, we suggest some approaches and methods for reducing their side-effects and clinical risk. We consider that using modern biotechnology to improve the tissue specificity to inflammatory site and tumor will be an important development direction of such biologics.
Biological Products ; metabolism ; Cytokines ; Humans ; Inflammation

Objective:To analyze the feasibility of hippocampal-avoidance (HA) prophylactic cranial irradiation (PCI) in small cell lung cancer patients (SCLC)(limited stage) after chemotherapy and thoracic radiation.Methods:From June 2016 to March 2019, 40 eligible SCLC patients were recruited and randomly divided into the routine PCI ( n=22) and hippocampal-avoidance PCI (HA-PCI) groups ( n=18). The HA zone was contoured according to the criteria of RTOG 0933. Volumetric-modulated arc therapy (VMAT) was adopted in the HA-PCI group. After radiotherapy, Hopkins verbal learning test (HVLT) and MRI were performed. Results:The average hippocampus volume was (4.01±1.57) cm 3, the average HA volume was (20.13±4.14) cm 3, HA D 100% was (7.19±0.38) Gy and HA D max was (14.38±1.18) Gy. During HVLT, 1-month-after-PCI vs. before-PCI (trial3, trial4, learning, percent retained), 1-month-after-PCI vs. after-PCI (trial3, learning), HA-PCI cohort showed advantages over PCI in HVLT scores. The average follow-up time was (17.00±8.47) months. Two patients with brain metastases which were out of the HAZ received routine PCI. Conclusions:PCI using VMAT technology to protect hippocampus is feasible in dosimetry. The test results indicate that the protective effect of hippocampus protection on memory is worthy of further promotion in clinical practice.

Linezolid is an antibacterial agent for the treatment of multi-resistant Gram-positive bacterial infections， which is widely used in clinical practice. However， there are large individual differences in the pharmacokinetic characteristics of the drug in patients， and it is difficult to obtain the optimal therapeutic effect when the drug is administered according to the conventional dose in the instructions. Therefore， it is necessary to carry out therapeutic drug monitoring （TDM） for linezolid， and guide and optimize its antibacterial treatment plan by using population pharmacokinetics （PPK） and pharmacodynamics principles. This paper summarizes the PPK changes and the research progress of individualized administration of linezolid in various populations， and recommends that the patient’s steady-state blood concentration is kept at 2-8 mg/mL through TDM when using linezolid clinically. It is recommended to appropriately reduce the dosage for patients with liver and kidney dysfunction， appropriately increase the dosage for obese， burned and children patients， and provide pharmaceutical monitoring during the medication process to promote rational drug use.