ABSTRACT
Objective:To prepare 99Tc m-hydrazinonicotinamide(HYNIC)-αCD8/Fab ( 99Tc m-αCD8/Fab), and explore the predictive value of 99Tc m-αCD8/Fab SPECT/CT imaging for the efficacy of anti-programmed death-1 (PD-1) immunotherapy. Methods:The αCD8/Fab was modified with HYNIC- N-hydroxysuccinimide (NHS) and IRDye800-NHS to form HYNIC-αCD8/Fab and IRDye800-αCD8/Fab (Dye-αCD8/Fab), respectively. 99Tc m-αCD8/Fab was prepared in sodium bicarbonate buffer (pH=8.5), with SnCl 2 being used as the reducing agent. The labeling yield and radiochemical purity of 99Tc m-αCD8/Fab and its stability in PBS and fetal bovine serum (FBS) were tested in vitro. The mouse spleen and human peripheral blood lymphocytes were isolated for cell-specific binding and blocking experiments of 99Tc m-αCD8/Fab in vitro. SPECT/CT imaging was used to analyze the specific binding ability of the 99Tc m-αCD8/Fab probe in CT26 colon cancer mouse models (BALB/c). The near infrared fluorescence imaging and SPECT/CT imaging were performed to detect the intra-tumoral CD8 + T cell infiltration after anti-PD-1 therapy in tumor bearing mice, and the results were further verified by HE and immunofluorescence staining. CD8 + T cell depletion study was performed to determine the role of CD8 + T cells in the tumor responses to anti-PD-1 therapy. Two-way analysis of variance was used to compare the data difference. Results:The labeling yield of 99Tc m-αCD8/Fab was 90% with a high radiochemical purity (95%) and good stability in vitro (radiochemical purity still more than 80% after 720 min in PBS and FBS). 99Tc m-αCD8/Fab could specifically bind to mouse CD8 + T cells ((10.30±0.81) percent added radioactive dose (%AD)/10 6 cells), compared with the binding ability in human peripheral blood lymphocytes group and CD8 antibody blocking group ((1.78±0.61) and (1.59±0.25) %AD/10 6 cells; F=10.07, P<0.001). SPECT/CT imaging showed that 99Tc m-αCD8/Fab had markedly higher tumor uptake in the CT26 colon cancer mouse models. Near-infrared fluorescence imaging showed that the tumor uptake of 99Tc m-αCD8/Fab in the responsive group was significantly higher than in the nonresponsive group after anti-PD-1 treatment ((8.9±1.1)% vs (7.1±0.8)%; F=4.69, P=0.024), and SPECT/CT imaging found the similar result. HE and immunofluorescence staining of tumor and lymph nodes showed that the proportion of lymphocyte infiltration was higher in the responsive group. Furthermore, CD8 + T cell depletion significantly reversed the therapeutic effect of anti-PD-1 immunotherapy in tumor-bearing mice. Conclusions:In this study, 99Tc m-αCD8/Fab was successfully obtained. CD8-specific SPECT imaging could sensitively visualize the tumor-infiltrating CD8 + T cells, suggesting the potential application value to predict and evaluate the efficacy of immunotherapy in the clinical settings.
ABSTRACT
OBJECTIVE@#To investigate the expression of CDC25A in non- small cell lung cancer (NSCLC) tissues and explore its correlation with the clinicpathological features of the patients and the expressions of let-7a1 and let-7c.@*METHODS@#We collected surgical specimens of pathologically confirmed NSCLC tissues and paired adjacent lung tissues from 44 patients and tissues of benign lung lesions from 9 patients. The expressions of CDC25A protein and mRNA in the tissues were detected by immunohistochemistry and fluorescence quantitative RT-PCR, respectively; the expressions of let-7a1 and let-7c mRNA were detected using tail-adding fluorescence quantitative RT-PCR.@*RESULTS@#The positivity rate of CDC25A protein expression was significantly higher in NSCLC tissues than in the adjacent tissues and benign pulmonary lesions (@*CONCLUSIONS@#The expression level of CDC25A is significantly increased in NSCLC with a negative correlation with Let-7c expression, which identifies CDC25A as a possible downstream target gene of Let-7c.
Subject(s)
Humans , Carcinoma, Non-Small-Cell Lung/genetics , Gene Expression Regulation, Neoplastic , Lung , Lung Neoplasms/genetics , Lymphatic Metastasis , MicroRNAs , RNA, Messenger/genetics , cdc25 PhosphatasesABSTRACT
Obejective To explore the diagnostic and curative evaluation value of gastrin-releasing pep-tide precursor (ProGRP) and neuron specific enolization enzyme (NSE) in small cell lung cancer (SCLC). Methods Sixty SCLC patients, sixty non-small cell lung cancer (NSCLC) patients and forty patients with be-nign pulmonary disease were collected fromJanuary 2014 to October 2015. The levels of serum ProGRP and NSE in all patients were determined by ELISA method and radioimmunoassay respectively then the clinical value of ProGRP and NSE on SCLC was evaluated. Results The levels of ProGRP and NSE in SCLC group were signif-icantly higher than those in NSCLC group and those in lung benign disease group (P 0.05). Conclusion ProGRP and NSE can be used as markers for the diagnostic and curative evaluation of SCLC. And ProGRP has higher sensitivity and specificity than NSE and can be promoted in clinic.
ABSTRACT
@#Objective To explore the effect of Drotaverine hydrochloride on preventing bladder spasm after transurethral prostatectomy.Methods 124 patients after transurethral prostatectomy were divided into patient-controlled epidural analgesia pump group (group I, n=61) and Drotaverine hydrochloride group (group II, n=63). Group I received bupivacaine by patient-controlled epidural analgesia, and the pump was withdrawed after 72 h. Group II received Drotaverine hydrochloride by intramuscular injection, 80 mg every 12 h, and then orally taken after anal exhaust for 3 days. Bladder spasm and adverse reaction were recorded in both groups. Results There was no significant difference in bladder spasm between group I (11.48%) and group II (12.70%) (P>0.05), as well as in side reaction between group I (16.39%) and group II (17.46%) (P>0.05). Conclusion Drotaverine hydrochloride is effective on preventing bladder spasm after transurethral resection of the prostate, with small side effect.