The CD19-targeting bispecific T-cell engager blinatumomab has shown remarkable efficacy in patients with relapsed/refractory B-cell precursor acute lymphoblastic leukemia. However, several studies showed that blinatumomab has a short plasma half-life due to its low molecular weight, and thus its clinical use is limited. Furthermore, multiple trials have shown that approximately 30% of blinatumomab-relapsed cases are characterized by CD19 negative leukemic cells. Here, we design and characterize two novel antibodies, A-319 and A-2019. Blinatumomab and A-319 are CD3/CD19 bispecific antibodies with different molecular sizes and structures, and A-2019 is a novel CD3/CD19/CD20 trispecific antibody with an additional anti-CD20 function. Our in vitro, ex vivo, and in vivo experiments demonstrated that A-319 and A-2019 are potent antitumor agents and capable of recruiting CD3 positive T cells, enhancing T-cell function, mediating B-cell depletion, and eventually inhibiting tumor growth in Raji xenograft models. The two molecules are complementary in terms of efficacy and specificity profile. The activity of A-319 demonstrated superior to that of A-2019, whereas A-2019 has an additional capability to target CD20 in cells missing CD19, suggesting its potential function against CD19 weak or negative CD20 positive leukemic cells.
Antigens, CD19/therapeutic use* ; Antineoplastic Agents/pharmacology* ; Humans ; Immunotherapy ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy* ; T-Lymphocytes

T-cell acute lymphoblastic leukemia (T-ALL) is one of the most dangerous hematological malignancies, with high tumor heterogeneity and poor prognosis. More than 60% of T-ALL patients carry NOTCH1 gene mutations, leading to abnormal expression of downstream target genes and aberrant activation of various signaling pathways. We found that chidamide, an HDAC inhibitor, exerts an antitumor effect on T-ALL cell lines and primary cells including an anti-NOTCH1 activity. In particular, chidamide inhibits the NOTCH1-MYC signaling axis by down-regulating the level of the intracellular form of NOTCH1 (NICD1) as well as MYC, partly through their ubiquitination and degradation by the proteasome pathway. We also report here the preliminary results of our clinical trial supporting that a treatment by chidamide reduces minimal residual disease (MRD) in patients and is well tolerated. Our results highlight the effectiveness and safety of chidamide in the treatment of T-ALL patients, including those with NOTCH1 mutations and open the way to a new therapeutic strategy for these patients.
Aminopyridines ; Benzamides ; Cell Line, Tumor ; Humans ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/metabolism* ; Proto-Oncogene Proteins c-myc/metabolism* ; Receptor, Notch1/metabolism* ; Signal Transduction ; T-Lymphocytes/metabolism*

Objective:To investigate the predictive value of the clinicopathological features of breast cancer for pathological complete response (pCR) after neoadjuvant chemotherapy (NAC) and to establish a predictive model based on the clinicopathological features.Methods:Clinicopathological data collected from 182 patients who underwent NAC and surgical treatment in Affiliated Hangzhou First People’s Hospital, Zhejiang University School of Medicine from Jan. 2013 to Dec. 2019 were retrospectively analyzed. The univariate and multivariate analysis were used to analyze the relationship between clinicopathological features and pCR after neoadjuvant chemotherapy. The predictive value in predicting the efficacy of NAC was evaluated, the receiver operating characteristic (ROC) curve and Nomogram prediction model were constructed.Results:Multivariate Logistic regression analysis showed that progesterone receptor (PR) , human epidermal growth factor 2 (HER2) and platelet distribution width (PDW) were independent predictors of pCR after NAC for breast cancer. The area under the curve (AUC) of model for predicting efficacy of NAC was 0.810 (95% CI:0.745-0.864) and the sensitivity and specificity was 68.75% and 82.67% respectively when the Jordan Index is at its maximum. Conclusion:ER-, HER2+ and PDW≤13.4% show better efficacy of NAC. The Nomogram model based on them can accurately predict the efficacy of NAC and can provide a reference for the selection of treatment options in clinical diagnosis and treatment.

Cashmere, also known as soft gold, is produced from the secondary hair follicles (SHFs) of cashmere goats. The number of SHFs determines the yield and quality of cashmere; therefore, it is of interest to investigate the transcriptional profiles present during cashmere goat hair follicle development. However, mechanisms underlying this development process remain largely unexplored, and studies regarding hair follicle development mostly use a murine research model. In this study, to provide a comprehensive understanding of cellular heterogeneity and cell fate decisions, single-cell RNA sequencing was performed on 19,705 single cells of the dorsal skin from cashmere goat fetuses at induction (embryonic day 60; E60), organogenesis (E90), and cytodifferentiation (E120) stages. For the first time, unsupervised clustering analysis identified 16 cell clusters, and their corresponding cell types were also characterized. Based on lineage inference, a detailed molecular landscape was revealed along the dermal and epidermal cell lineage developmental pathways. Notably, our current data also confirmed the heterogeneity of dermal papillae from different hair follicle types, which was further validated by immunofluorescence analysis. The current study identifies different biomarkers during cashmere goat hair follicle development and has implications for cashmere goat breeding in the future.

Objective:To investigate the efficacy and clinical experiences of fluorescein angiography assisted occlusion via lateral-orbital keyhole approach in internal carotid bifurcation aneurysms. Methods:The clinical data of 16 patients with internal carotid artery bifurcation aneurysms admitted to our hospital from January 2016 to December 2020 were retrospectively analyzed. All patients accepted fluorescein angiography assisted occlusion via lateral-orbital keyhole approach. The patients were followed up at one, 3, and 6 months after surgery by medical imaging. The therapeutic efficacy of these patients was assessed by Glasgow outcome scale (GOS). Results:All aneurysms in these 16 patients were clipped at one-stage operation. Intraoperative fluorescein angiography and FLOW 800 showed that the aneurysms were completely clipped without residual, and the blood flow of the parent artery and perforating arteries was unobstructed. Six months after surgery, 14 patients recovered well (GOS scores of 5), and 3 developed limb hemiplegia (GOS scores of 4).Conclusion:Intraoperative fluorescein angiography assisted occlusion via lateral-orbital keyhole approach is safe and effective in internal carotid bifurcation aneurysms.

Objective:To investigate the value of CT enhancement degree in diagnosing cervical lymph node metastasis of papillary thyroid carcinoma (PTC) .Methods:CT data of 535 neck lymph nodes with pathological diagnosis in 251 cases were retrospectively analyzed. The ratio and difference between the density of CT enhancement and plain scan were calculated. Mann-Whitney test was used to analyze the distribution of ratio and difference in lymph node metastasis group and non-metastasis group. The optimal thresholds of ratio and difference in the two groups were obtained by receiver operating characteristic (ROC) curve analysis.Results:535 cervical lymph nodes including 271 in metastatic group and 264 in non-metastatic group. The ratios of two groups were 2.30 (2.04, 2.76) and 1.66 (1.51, 1.81) ( Z=-16.94, P<0.05) respectively, and the differences were 58 (49, 76) Hu and 31 (22, 36) Hu ( Z=-18.045, P<0.05) respectively. When the ROC area under curve of ratio and difference between the two groups were 0.923 and 0.951 respectively, the optimal thresholds were 1.93 and 39.5Hu respectively. The sensitivity and specificity of diagnosing lymph node metastasis were 84.9% and 87.1%、91.5% and 86.0%respectively.The sensitivity and specificity of combination of the two diagnosing lymph node metastasis were 82.3% and 91.7%. Conclusions:The ratio of enhancement and plain scan density ≥1.93 and the difference ≥39.5Hu have a high diagnostic efficiency for cervical lymph node metastasis of PTC. The specificity of the two is similar, but the latter has higher sensitivity. The combination of the two can significantly improve the specificity, thereby reducing unnecessary surgical trauma.

Objective To investigate the relationship between Hashimoto thyroiditis (HT) and thyroid papillary carcinoma (PTC) by analyzing the expression of BRAF V600E mutation and (N-,H-,K-) RAS codons 12,13 and 61 mutants in cases of multifocal PTC with HT.Methods 80 tumor samples in 37 multifocal PTC with HT cases,were analyzed for the genotypic changes of BRAF V600E,as well as the (N-,H-,K-)RAS codons 12,13 and 61 mutants by DNA sequencing assay and amplification refractory mutation system (ARMS).Results BRAF V600E mutation was detected in 51 samples and RAS gene mutations was found in 3 samples (N-RAS codon 61 mutant in 2 samples and H-RAS codon 61 mutant in 1 sample).Different clonal origin was present in 20 cases of multifocal PTC with HT (54.1％,20/37).There was no statistical significance (P ＞ 0.05) in the incidence of the difference in the origin of tumor cells,compared with the results (61.7％,37/60) of multifocal PTC without HT in the related literature.Conclusion In more than half of multifocal PTC with HT cases,the tumor cells originate from different clones.Our results do not support the opinion that HT predisposes patients to develop PTC,because HT does not have a significant effect on expression of BRAF and RAS gene mutation in PTC,accordingly HT is more likely to be a part of the host tumor immune response system.

Objective To investigate the hemodynamic monitoring and clinical significance of transcranial Doppler (TCD) combined with indocyanine green (ICG) fluorescence angiography and FLOW 800 in carotid endarterectomy (CEA).Methods Forty-eight patients with extracranial carotid atherosclerotic stenosis underwent CEA in our hospital from October 2015 to September 2018 were chosen to our study. The mean blood flow velocity (Vm) of the middle cerebral artery (MCA) was monitored by TCD throughout the operation. The intraoperative blood pressures were adjusted according to the monitoring results, and the average blood Vm of the narrowed arteries was measured using TCD probe before and after artery temporary occlusion. ICG fluorescence angiography and FLOW 800 were used to monitor vascular morphology and patency simultaneously.Results One patient, who was observed to have severe stenosis by TCD, ICG fluorescence angiography, and FLOW 800 after arteries temporary occlusion, showed significantly improved by multimodal monitoring after re-suture; one patient showed external carotid artery occlusion by ICG fluorescence angiography and FLOW 800 had noexternal carotid artery occlusion; other 46 patients showed no carotid artery stenosis and good intracranial perfusion by TCD, ICG fluorescence angiography, and FLOW 800 after initial suture. Two patients developed ischemic stroke (IS) after CEA, and both of them were recovered after conservative medical treatment for two weeks without any neurological dysfunction; two patients had cerebral hyperperfusion syndrome (CHS), and both of them relieved after blood pressure control three d after treatment; one patient had subcutaneous hematoma in the operation area, and the hematoma was gradually absorbed after conservative treatment; the rest patients recovered well. All patients were followed up for 4-6 months after CEA, and CT angiography showed that carotid artery stenosis was relieved; CT perfusion imaging and TCD examinations showed that cerebral perfusion was significantly improved as compared with those before operation.Conclusion Intraoperative TCD combined with ICG fluorescence angiography and FLOW 800 can effectively monitor the intracranial and extracranial blood flow of intraoperative carotid artery, which is of great significance in reducing the risk of IS and CHS during carotid endarterectomy.

Objective To compare the perioperative and recent efficacy of different mitral valve treatment strategies for mitral regurgitation caused by infective endocarditis. Methods From January 2009 to December 2014, 168 patients with mitral regurgitation caused by infective endocarditis underwent surgical treatment. Among them, mitral valve repair was performed in 65 patients (group A) and mitral valve replacement was performed in 103 patients (group B). Preoperative clinical profiles, perioperative details and follow-up data were reviewed retrospectively. Results There was no operative death in both groups. The cardiopulmonary bypass time and aorta clamp time were similar between the two groups. Intensive care stay was significantly shorter in group A [(1.4 ± 0.6) d] compared with group B [(1.9 ± 0.9) d](P＜0.05), and hospital stay had no significant difference between two groups (P＞0.05). Perioperative cerebral hemorrhage was observed in no patient in group A and 2 patients (1.9%) in group B. There was no in-hospital death in group A, and 2 patients (1.9%) in-hospital deaths occurred in group B. The difference was not statistically significant (P＞0.05). The incidence of heart-related adverse events in group A and group B was 6.2%(4/65) and 7.9%(8/101), and the incidence of infection related events was 0 and 3.0% (3/101) 3 years after operation (P＞0.05). Conclusions Mitral valve repair for patients with mitral regurgitation caused by infective endocarditis is safe and reliable, and the short-term effect is satisfactory. However, infection tissue should be eliminated thoroughly, and the related complications of valve replacement can be avoided and the risk of anticoagulation can be reduced.

Objective To investigate diagnostic value of CT signs in papillary thyroid carcinoma (PTC) by comparing CT signs of PTC with different sizes.Methods CT signs of 406 PTC from 396 patients confirmed by histology were analyzed retrospectively.Based on the largest tumor diameter,PTC were divided into 1.1-2.0 cm group,2.1-3.0 cm group and＞3.0 cm group.Distribution of irregular shape,cookie bite sign,enhanced narrow/ fuzzy and microcalcification in each group was analyzed.Results There were 318 pieces in 1.1-2.0 cm group,60 pieces in 2.1-3.0 cm group and 28 pieces in ＞3.0 cm group,respectively.The rate of irregular shape was 89.6％(285/318),75.0％(45/60) and 64.3％(18/28) in each group,the rate of cookie bite sign was 83.6％(266/318),71.7％(43/60) and 64.3％ (18/28),and enhanced narrow/fuzzy was 84％(267/318),78.3％ (47/60) and 67.9％ (19/28) in each group,and microcalcification was 35.5％ (113/318),40.0％ (24/60) and 59.3％ (16/27) in each group.The rate of irregular shape (x2=20.092,P=0.000)and cookie bite sign (x2=9.695,P=0.008)had statistical difference among the three groups,while the rate of enhanced narrow/fuzzy(x2=5.175,P=0.075)and microcalcification (x2=5.277,P=0.071) had no statistical difference among each group.Furthermore,irregular shape and cookie bite sign were compared between groups.Rate of irregular shape in 1.1-2.0 cm group and 2.1-3.0 cm (x2=9.746,P=0.002)group,1.1-2.0 cm group and ＞ 3.0 cm group (x2=15.180,P=0.000) was statistically different.Rate of cookie bite sign in 1.1-2.0 cm group and ＞3.0 cm(x2=6.560,P=-0.010)was statistically different.There was no statistical difference between other groups.Conclusions Although irregular shape,cookie bite sign,enhanced narrow/fuzzy and microcalcification are important CT signs in diagnosing PTC,distribution of different signs varies with tumor sizes.Correct identification of these differences will help to improve the accuracy of preoperative diagnosis,and reduce occurrence of misdiagnosis.