RÉSUMÉ
Objective:To prepare the anti-programmed death-ligand 1 (PD-L1) nanoantibody P3C8-C3Fab by ligating with C3Fab and to investigate its role in plasma half-life.Methods:The C3Fab peptide derived from protein G was molecularly fused with the nanobody P3C8 by DNA recombination technology. The nanoantibody P3C8-C3Fab was inducibly expressed and purified in the E. coli BL21 strain, and the binding of it to PD-L1 protein, mouse IgG, and PD-L1-expressing tumor cells was detected by enzyme-linked immunosorbent assay (ELISA). The residual P3C8-C3Fab was detected in mouse serum at different times using double-antibody sandwich ELISA to assess the prolongation of the plasma half-life of PD-L1 nanobodies by C3Fab. Results:The nanoantibody P3C8-C3Fab was successfully constructed, and it could efficiently express itself in soluble form in BL21. The purified NbP3C8-C3Fab protein was obtained with a mass fraction of about 90% at a yield of 7.18 mg/L. The affinity of P3C8-C3Fab for PD-L1 protein and mouse IgG gradually increased with increasing mass concentration and showed a concentration correlation. The binding of P3C8-C3Fab to lung cancer A549 cells showed a concentration correlation. The concentration standard curve of P3C8-C3Fab in mouse serum showed a typical S-shape with a concentration correlation. The plasma half-life of P3C8 was only 0.44 h, while the plasma half-life of P3C8-C3Fab was 21.27-fold higher, up to 9.36 h.Conclusions:The linkage of C3Fab to the nanobodies of P3C8 can significantly prolong the plasma half-life of P3C8, which is valuable for the improvement of in vivo nanobody effects.
RÉSUMÉ
The co-existence of multiple chronic diseases has been increasing in the elderly population, it has become a major challenge globally, and identifying comorbidities patterns can help provide clues for disease prevention and treatment, as well as improving prognosis. This article reviews the identification methods, influencing factors and management strategies of chronic disease comorbidities, to provide a reference for the research and management of comorbidity.
RÉSUMÉ
Objective:To established a method for the detection of soluble programmed death ligand 1 (PD-L1) protein in serum based on the poly nanoantibody of lumazine synthase(LS).Methods:A dual nanobody-based sandwich ELISA was established with a competitive ELISA to screen nanobodies recognizing different epitopes of PD-L1 as paired antibodies. To improve sensitivity, PD-L1 nanobody P3C8 and lumazine synthase(LS) were fused, and nanobodies were obtained in polymeric forms as sPD-L1 protein captures, so as to develop an LS-displayed polymeric nanobody-based sandwich ELISA (LSNbs-ELISA) method to detect sPD-L1.Results:Compared with the Nbs-ELISA method, the LSNbs-ELISA method is approximately 11-fold more sensitive for sPD-L1 detection. The limit of detections (LODs) of Nbs-ELISA and LSNbs-ELISA for sPD-L1 in serum were 2.87 ng/ml and 0.255 ng/ml, respectively. Both assays were highly specific for the detection of sPD-L1 and did not react with structure-related proteins PD-1, CD27, CD70, CD137, and CD147 when spiked into the human serum.Conclusions:The Nbs-ELISA and LSNbs-ELISA assays both have high sensitivity and specificity for detecting sPD-L1 in serum and could have potential clinical applications.
RÉSUMÉ
Aim To investigate TY501′s role in bleo-mycin ( BLM )-induced pulmonary fibrosis in rats. Methods Forty rats were randomly divided into 5 groups, including the sham operation, BLM, PFD, TY501(high and low dose) groups. After administra-tion of BLM intratracheally, PFD and TY501 were giv-en in each group daily, according to the dosage de-signed during 21 days. Lung coefficient, PaO2 were tested before killing the rats. The contents of ALB, ALP, LDH, GSH, HYP were detected by regent kit respectively. PCⅢ and COL4 were determined by ELISA. Results ( 1 ) Some indicators of alveolitis in early stage of IPF: the contents of lung coefficient in three treatment groups were lower and PaO2 was higher than those in BLM group ( P<0. 05 ); compared with BLM group, the contents of ALB, ALP, LDH in the treatment groups reduced on 21 st day ( P<0. 05 );the expression of GSH in BLM group was increased for feedback regulation and higher than the treatment groups and the sham operation group (P<0. 05);(2) some indicators of pulmonary interstitial fibrosis in late stage of IPF:the expressions of HYP, PCⅢand COL4 were reduced after the treatment. There were signifi-cant differences compared with BLM group ( P <0. 05 ) . Conclusions TY501 is valuable for the ther-apy of IPF, the same as the positive drug pirfenidone. TY501 attenuates BLM-induced pulmonary fibrosis, which may be related to the affection of TGF-βpathway and inhibition of MMPs.
RÉSUMÉ
Objective To investigate the MR characteristics of ulnar nerve and muscles in cubital tunnel syndrome.Methods Twenty eight patients with cubital tunnel syndrome and 28 asymptomatic volunteers underwent MR imaging,MR neurography was performed by using an isotropic three dimensions T2 sampling perfection with application optimized contrasts using different flip angle evolutions (3D T2 SPACE)on 15 patients with cubital tunnel syndrome.To evaluate changes in T2 signal intensity of the ulnar nerve,regions of interest were placed in the center of the location of the highest apparent ulnar nerve signal intensity on the axial FS T2WI images and in normal muscle within the same image slice,and the ratio of signal intensity was calculated.The sensitivity of 3D T2 SPACE sequence in detecting cubital tunnel syndrome was determined.The standard t tests were used to assess whether ulnar nerve size and relative signal intensity in symptomatic patients were statistically different from normal volunteers.Results The cross-sectional area of ulnar nerve in 24 patients and 2 volunteers was enlarged,the signal intensity of ulnar nerve in 26 patients and 16 volunteers was increased.Increased signal and muscle atrophy adjacent to the ulnar nerve were detected in 4 patients.Cubital tunnel syndrome was detected in 14 patientson 3D T2 SPACE sequence.The mean ulnar nerve sizes in the symptomatic and normal group were (0.15±0.06)and (0.06±0.01)cm2 respectively,the mean relative signal intensities in the symptomatic and normal groups were (2.86± 1.45) and 1.57±0.39 respectively (t values were 2.220 and 4.546,P＜0.05).Conclusions Ulnar nerve size and T2 signal intensity were increased,in patients with cubital tunnel syndrome.In addition,muscles innervated by the ulnar nerve showed atrophy with increased T2 signal intensity.
RÉSUMÉ
Objective To investigate the site,MR signal types and morphological characteristics of ulnar nerve in cubital tunnel for healthy adults,in order to improve the awareness of ulnar nerve.Methods Unilateral elbow of forty healthy volunteers were scanned with MR,the sequences were as follows:T1-weighted-spin-echo,T2-weighted-spin-echo-fat-suppression,PD-weighted-spin-echo-fat-suppression,among 40,13 were supine,27 were prone.The site and MR signal types and morphological characteristics of ulnar nerve were observed,the long diameter and short diameter of the ulnar nerve on different axials were respectively measured.Results On axial,ulnar nerve was posterior to the medial condyle of the humerus at proximal elbow,lied between the flexor carpi ulnaris and the flexor digitorum profunds and superficialis muscles distally.The shape of the ulnar nerve was roundness or elliptic,the signal of 40 volunteers'(100％) ulnar nerve was isointensity on T1-weighted,the signal of 17 volunteers'(42.5％)ulnar nerve was isointense on T2-weighted or PD-weighted,the MRI signal of 23 volunteers' (57.5％) ulnar nerve was slight hyperintense on T2-weighted or PD-weighted,especially on the axial of medial condyle of humerus.The variation range of long diameter and short diameter of the ulnar nerve respectively were 1.4-3.8 mm,1.0-3.0 mm.Conclusion The certain location,MR signal types and morphological characteristics of normal ulnar nerve play an important role in the diagnosis of cubital tunnel syndrome.
RÉSUMÉ
The purpose of the study is to construct recombinant goat pox virus (GPV) expressing Peste des petits ruminants virus (PPRV) H protein, and to evaluate the immunization effect. Recombinant GPV containing PPRV H gene (rGPV-PPRV-H) was selected and purified by gpt and eGFP utilizing plaque purification, and the final selected recombinant GPV was proved to be purified by PCR. Immunofluorescence and Western blotting showed that the recombinant virus could express H protein of PPRV while infecting lamb testis cells. Six goats were immunized with 2 x 10(6) PFU rGPV-PPRV-H through intradermal injection, and were immunized for the second time at 28 days with the same dose recombinant virus after first immunization. Serum was collected after immunization, and was analyzed for the neutralization antibodies. 21 days after first immunization, the neutralization antibodies of GPV were 40, 80, > or = 80, > or = 80, 40, > or = 80 in turn, and neutralization antibodies of PPRV were 80, 80, 80, 80, 40, 40, 10 in turn; 14 days after second immunization, the neutralization antibodies of GPV were all > or = 80, and the neutralization antibodies of PPRV were > 80, 80, > 80, 80, 80 and 40 in turn. This study established a foundation for the industrialization of the PPRV recombinant GPV vaccine.