Preparation of mouse monoclonal antibodies against human adenovirus 55 Hexon (HAdV55 Hexon) protein / 细胞与分子免疫学杂志

Objective To prepare specific mouse monoclonal antibody (mAb) against human adenovirus type 55 Hexon protein (HAdV55 Hexon). Methods The Hexon genes of HAdV55, 3, 4, 7, 16 and 21 were chemically synthesized as templates for PCR amplification. The prokaryotic expression plasmids pET28a-HAdV55 Hexon and eukaryotic expression plasmids pCAGGS-HAdV3, 4, 7, 16, 21 and 55 Hexon were constructed respectively. The pET28a-HAdV55 Hexon plasmid was transformed into E. coli competent cell BL21 (DE3) and was induced by IPTG. After the purified inclusion body was denatured and renatured, Hexon55 protein was purified by tangential flow filtration system. pCAGGS-HAdV55 Hexon was used to immunize BALB/c mice by cupping, and HAdV55 Hexon protein was used to booster immunization. The anti-HAdV55 Hexon mAb was prepared by hybridoma technique and the titer and subclass were determined. The specificity of antibody was identified by Western blot using HEK293T cells transfected with pCAGGS-HAdV55 Hexon and by immunofluorescence assay (IFA) using BHK cells transfected with pCAGGS-HAdV55 Hexon. Both clones with high titer were selected, and the cross-reactivity of pCAGGS-HAdV3, 4, 7, 16, 21 and 55 Hexon transfected cells were analyzed by Western blot analysis and IFA. Results PET28a-HAdV55 Hexon and pCAGGS-HAdV55 Hexon, 3, 4, 7, 16 and 21 expression plasmids were successfully constructed. BL21 transformed with pET28a-HAdV55 Hexon was induced by IPTG. The HAdV55 Hexon protein was mainly expressed in the form of inclusion body. After denaturation and renaturation, the purified HAdV55 Hexon protein was obtained by ultrafiltration. Six hybridoma cell lines secreting HAdV55 Hexon mAb were obtained. The antibody subclass analysis showed that 2 strains were IgG2a subtypes and 4 strains were IgG2b. Two specific HAdV55 Hexon antibodies with high titer were obtained, and there was no cross-reactivity with HAdV3, 4, 7, 16, 21 Hexon. Conclusion The specific mice mAb against HAdV55 Hexon provides an experimental basis for establishing its antigen detection method.

Safety and effectiveness of total hip arthroplasty in patients with hypothyroidism / 中国修复重建外科杂志

Objective: To evaluate the safety and effectiveness of total hip arthroplasty (THA) in patients with hypothyroidism. Methods: Sixty-three patients with hypothyroidism (hypothyroidism group) and 63 euthyroid patients without history of thyroid disease (control group) who underwent primary unilateral THA between November 2009 and November 2018 were enrolled in this retrospective case control study. There was no significant difference between the two groups in gender, age, body mass index, hip side, reason for THA, American Society of Anesthesiology (ASA) classification, preoperative hemoglobin (Hb) level, and preoperative Harris score ( P>0.05). The perioperative thyroid stimulating hormone (TSH) and thyroxine (T 4) levels, the hypothyroidism-related and other complications during hospitalization, the decrease in Hb, perioperative total blood loss, blood transfusion rate, length of hospital stays, and 90 days readmissions rate in the two groups were recorded and evaluated. The periprosthetic joint infection, aseptic loosening of the prosthesis, and hip Harris score during follow-up were recorded. Results: The differences in the TSH and T 4 of hypothyroidism group between pre- and 3 days post-operation were significant ( P>0.05) and no hypothyroidism-related complications occurred after THA. The decrease in Hb and perioperative total blood loss in the hypothyroidism group were significantly higher than those in the control group ( P0.05). No significant difference in the rate of complications (liver dysfunction, heart failure, pulmonary infection, urinary infection, and wound complication) between the two groups was found ( P>0.05) except for the rate of intramuscular vein thrombosis which was significantly lower in the hypothyroidism group, and the rate of postoperative anemia which was significantly higher in the hypothyroidism group ( P<0.05). The two groups were followed up 1.0-9.9 years (mean, 6.5 years). At last follow-up, Harris score in both groups were significantly higher than those before operation ( P<0.05). An increase of 39.5±12.3 in hypothyroidism group and 41.3±9.3 in control group were recorded, but no significant difference was found between the two groups ( t=0.958, P=0.340). During the follow-up, 1 case of periprosthetic joint infection occurred in the hypothyroidism group, no loosening or revision was found in the control group. Conclusion: With the serum TSH controlled within 0.5-3.0 mU/L and T 4 at normal level preoperatively, as well as the application of multiple blood management, hypothyroid patients can safely go through THA perioperative period and effectively improve joint function, quality of life, and obtain good mid-term effectiveness.