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The presence of HLA antibodies is widely recognized as a barrier to solid organ transplantation, and for lung transplant candidates, it has a significant negative impact on both waiting time and waiting list mortality. Although HLA antibodies have been associated with a broad spectrum of allograft damage, precise characterization of these antibodies in allosensitized candidates may enhance their accessibility to transplant. The introduction of Luminex-based single antigen bead (SAB) assays has significantly improved antibody detection sensitivity and specificity, but SAB alone is not sufficient for risk-stratification. Functional characterization of donor-specific antibodies (DSA) is paramount to increase donor accessibility for allosensitized lung candidates. We describe here our approach to evaluate sensitized lung transplant candidates. By employing state-of-the-art technologies to assess histocompatibility and determine physiological properties of circulating HLA antibodies, we can provide our Clinical Team a better risk assessment for lung transplant candidates and facilitate a “road map” to transplant. The cases presented in this paper illustrate the “individualized steps” taken to determine calculated panel reactive antibodies (cPRA), titer and complement-fixing properties of each HLA antibody present in circulation. When a donor is considered, we can better predict the risk associated with potentially crossing HLA antibodies, thereby allowing the Clinical Team to approach allosensitized lung patients with an individualized medicine approach. To facilitate safe access of sensitized lung transplant candidates to potential donors, a synergy between the histocompatibility laboratory and the Clinical Team is essential. Ultimately, donor acceptance is a decision based on several parameters, leading to a risk-stratification unique for each patient.

The impact of COVID‐19 vaccination on the alloimmunity of transplant candidates is unknown. We report a case of positive B cell flow cytometry crossmatch in a patient waiting for second kidney transplantation, 37 days after receiving the COVID‐19 vaccine. The preliminary crossmatch, using sample collected before COVID‐19 vaccination, was negative. The antibodies to mismatched donor HLA‐DR7 were detected only with multi‐antigen beads but not with single‐antigen beads, excluding possible prozone effects in solid‐phase antibody assays. The crossmatches were positive with HLA‐DR7–positive surrogates (n = 2) while negative with HLA‐DR7–negative surrogates (n = 3), which confirms the HLA‐DR7 alloreactivity. The antigen configurations on B lymphocytes are similar to that on the multi‐antigen beads while distinct from the single‐antigen beads. HLA‐DR7 was the repeating mismatched antigen with the failing first kidney allograft. The newly emerged antibody to HLA‐DR7 probably is the consequence of bystander activation of memory response by the COVID‐19 vaccination. This case highlights the importance of verifying allo‐sensitization history and utilizing multiple assays, including cell‐based crossmatch and solid‐phase assays with multi‐antigens. COVID‐19 immunization may deserve special attention when assessing the immunological risk before and after organ transplantation.

Background The risk of locoregional recurrence (LRR) after mastectomy for breast invasive micropapillary carcinoma (IMPC) remains poorly defined. We aimed to construct an effective prognostic nomogram to estimate the individualized risk of LRR for providing accurate information for long-term follow-up. Patients and Methods A total of 388 patients with breast IMPC were included in the current study. Based on the Cox regression and clinical significance, a nomogram with an online prediction version was created. This model was evaluated and internally validated by concordance index and calibration plot. Receiver operating characteristic curve and decision curve analysis were used to assess the discrimination and clinical utility, and Kaplan-Meier curves estimated the probability of LRR. Results The variables (age, lymph node metastasis, hormone receptor status, lymphovascular invasion, histologic grade, and adjuvant radiotherapy) were included in the nomogram. This model was well-calibrated to predict the possibility of LRR and displayed favorable clinical utility; the concordance index was 0.86 (95% confidence interval, 0.81-0.91), which was higher than any single predictor. The area under the curve of the nomogram was 0.89, whereas that of the conventional staging system was 0.72. An online prognostic nomogram was built for convenient use. Kaplan-Meier curves showed that the nomogram had a better risk stratification than the conventional staging system. Conclusions The nomogram could accurately predict the individualized risk of LRR after mastectomy for breast IMPC. By identifying the risk stratification, this model is expected to assist clinicians and patients in improving long-term follow-up strategies.

PurposeEnhanced B-cell presentation of donor alloantigen relative to presentation of HLA-mismatched reference alloantigen is associated with acute cellular rejection (ACR), when expressed as a ratio called the antigen presenting index (API) in an exploratory cohort of liver and intestine transplant (LT, IT) recipients.MethodsTo test clinical performance, we measured the API using the previously described 6-hour assay in 84 LT and 54 IT with median age 3.3 years (0.05-23.96). Recipients experiencing ACR within 60 days after testing were termed rejectors.ResultsWe first confirmed that B-cell uptake and presentation of alloantigen induced and thus reflected the alloresponse of T-helper cells, which were incubated without and with cytochalasin and primaquine to inhibit antigen uptake and presentation, respectively. Transplant recipients included 76 males and 62 females. Rejectors were tested at median 3.6 days before diagnosis. The API was higher among rejectors compared with non-rejectors (2.2 ± 0.2 vs 0.6 ± 0.04, p-value=1.7E-09). In logistic regression and ROC analysis, API ≥ 1.1 achieved sensitivity, specificity, positive and negative predictive values for predicting ACR in 99 training set samples. Corresponding metrics ranged from 80-88% in 32 independent post-transplant samples, and 73-100% in 20 independent pre-transplant samples. In time-to-event analysis, API ≥ 1,1 predicted higher incidence of late DSA after API measurements in LT (p=0.011) and graft loss in IT recipients (p=0.008), compared with recipients with APIConclusionEnhanced donor antigen presentation by circulating B-cells predicts rejection after liver or intestine transplantation as well as higher incidence of DSA and graft loss late after transplantation

Virtual crossmatch (VXM) is used as an alternative to or in conjunction with a cell-based physical crossmatch (PXM) for assessing HLA (human leukocyte antigen) compatibility prior to deceased donor kidney transplantation (DDKT). Data on practice patterns and perceptions regarding VXM use in the US are limited. We performed a survey of US HLA directors and transplant surgeons regarding HLA testing and crossmatch strategies. 53 (56 %) HLA directors and 68 surgeons (representing ∼ 23 % of US transplant centers) completed the survey. Both groups agreed that VXM could reduce cold ischemia time (CIT), costs and improve allocation efficiency. VXM use increased following the 2021 kidney allocation change. Reducing CIT was the primary reason for favoring VXM over PXM. Preference for VXM reduced as candidates' panel reactive antibodies increased. Regulations, program policies and limitations of HLA technology were cited as important reasons for preferring PXM over VXM. Surgeons reported similar perceptions, but findings are limited by the low response rate. Finally, half the labs reported lacking specific protocols for VXM use. In conclusion, improved HLA technology and protocols along with changes to institutional procedures and policy regulations are needed for safer expansion of VXM in DDKT.

Background Allosensitization in heart transplant candidates is associated with longer transplant wait times and post-transplant complications. We summarize our experience with desensitization using carfilzomib , an irreversible proteasome inhibitor that causes plasma cell apoptosis . Methods One cycle of desensitization consisted of plasmapheresis and carfilzomib 20 mg/m 2 on days 1, 2, 8, 9, 15, and 16 with intravenous immune globulin 2 g/kg after carfilzomib on day 16. Patients underwent repeat cycles as indicated. We compare calculated panel-reactive antibody (cPRA) for neat combined Class I and II IgG and C1q pre- and post-treatment using a cutoff for cPRA entry of ≥ 4000 and 500 MFI, respectively. Results From June 2013 to October 2019, 9 patients underwent 20 cycles of carfilzomib-based desensitization. Each cycle resulted in an average cPRA decrease of 24% (95% CI: 6-42) for IgG and 36% (95% CI: 17-55) for C1q. From treatment start to finish, mean cPRA fell from 76% to 40% (p = 0.01) for IgG and 56% to 4% (p = 0.017) for C1q. Six of 9 patients have been transplanted with 5 of the transplanted hearts crossing preoperative donor-specific antibodies. During a median follow-up of 35.1 months, all transplanted patients have survived with only 1 occurrence of treated rejection. Side effects of desensitization included acute kidney injury (67%) and thrombocytopenia (33%) with all episodes self-resolving. Conclusions A carfilzomib-based desensitization strategy among heart transplant candidates reduces the level of HLA antibodies and complement binding , facilitates successful transplantation, and is associated with excellent outcomes at 3 years.

Chronic lung allograft dysfunction (CLAD) is the major complication limiting long-term survival in lung transplant recipients (LTRs), with those developing donor-specific anti-human leukocyte antigen (HLA) antibodies (DSAs) previously found to have increased risk for CLAD. However, as DSA responses vary in timing of development, specificity, breadth, persistence, and complement-binding capacity, we hypothesized that these characteristics would impact CLAD and survival outcomes.We retrospectively analyzed DSA characteristics and outcomes in a single-center cohort of 582 LTRs who had serum samples collected prospectively from 2010 to 2016. Luminex-based single antigen bead assays were performed to assess DSA.DSAs were detected in 247 LTRs (42%), of which 124 (21.3%) were de novo DSAs and 53 (9.1%) were complement-binding (C1q+). CLAD developed in 208 LTRs (35.7%) during the follow-up period, with 67.8% determined as bronchiolitis obliterans syndrome phenotype and 32.2% as restrictive allograft syndrome phenotype. We found a shorter time to CLAD in LTRs with persistent DSAs (p = 0.04) and HLA-DQ-specific DSAs (p = 0.03). LTRs who developed C1q+ DSAs had significantly shorter time to CLAD (p0.001), with 100% of C1q+ DSAs being persistent and no differences between CLAD phenotypes. CLAD-free survival was significantly reduced in LTRs who developed C1q+ DSAs (p = 0.001), HLA-DQ-specific DSAs (p = 0.03), and multiple DSAs (p = 0.02).Together, our findings demonstrate that DSA characteristics of persistence, HLA-DQ specificity, and C1q+ DSAs are associated with shorter time to CLAD. Additionally, C1q+, HLA-DQ-specific, and multiple DSAs are associated with decreased CLAD-free survival. These characteristics may improve DSA risk stratification for deleterious outcomes in LTRs.

Radiation resistance affects survival in non-small-cell lung cancer (NSCLC) patients. Further exploration of mechanisms and targets is urgently needed. Using bioinformatic analyses, we found that UBE2T is associated with survival, tumor size, lymph node metastasis and distant metastasis. Then, real-time PCR and immunohistochemistry were performed to explore the differentially expressed genes between normal and NSCLC tissues. Furthermore, we used colony formation, EdU incorporation, scratch, transwell assays, flow cytometry, immunofluorescence and western blot to assess the role of UBE2T in vitro and in vivo. RNA-Seq and coimmunoprecipitation were used to explore the mechanism. The results showed that UBE2T promotes proliferation, migration, invasion, and radiation resistance in vitro and in vivo by accelerating the G2/M transition and inhibiting apoptosis. Mechanistically, UBE2T promotes epithelial-mesenchymal transition (EMT) via ubiquitination-mediated FOXO1 degradation and Wnt/β-catenin signaling pathway activation. Moreover, FOXO1 reversed radiation resistance and EMT. Therefore, UBE2T may be a potential target for enhancing radiotherapy sensitivity and serve as a biomarker to predict prognosis.

Tumor necrosis factor alpha-induced protein 8 (TNFAIP8) is implicated in the tumor progression and prognosis of triple-negative breast cancer (TNBC), but the detailed regulatory mechanism of TNFAIP8 in cisplatin tolerance in TNBC has not yet been investigated. TNFAIP8 was evidently upregulated in TNBC tumor tissues and cell lines. Knocking down TNFAIP8 led to impaired proliferation and elevated apoptosis of TNBC cells upon cisplatin (DDP) treatment. Mechanistic studies revealed that TNFAIP8 repressed the expression of p53 and p53-promoted microRNA (miR)-205-5p; moreover, miR-205-5p targeted multiple genes required for the cell cycle and repressed Akt phosphorylation, which thus inhibited the proliferation of TNBC cells. In addition, silencing of TNFAIP8 led to the upregulation of miR-205-5p and the restraint of the TRAF2-NF-κB pathway, which thus enhanced the suppressive effects of DDP on tumor growth in nude mice. This study revealed that TNFAIP8 was essential in the DDP tolerance formation of TNBC cells by reducing p53-promoted miR-205-5p expression. Thus, targeting TNFAIP8 might become a promising strategy to suppress TNBC progression., Graphical Abstract, In the present study, we discovered that TNFAIP8 was essential in the DDP tolerance formation of TNBC cells by reducing p53-promoted miR-205-5p expression. Thus, targeting TNFAIP8 might become a promising strategy to suppress TNBC progression.

Background Long non-coding RNA H19 (lncRNA H19) has been implicated in tumorigenesis and metastasis of breast cancer through regulating epithelial to mesenchymal transition (EMT); however, the underlying mechanisms remain elusive. Methods LncRNA H19 and TNFAIP8 were identified by qRT-PCR and western blotting. CCK-8 assay, clone formation assay, transwell assay, and flow cytometry assay were performed to determine cell proliferation, migration, invasion and cell cycle of breast cancer respectively. Western blotting and immunohistochemistry (IHC) were utilized to evaluate the protein expression levels of p53, TNFAIP8, and marker proteins of EMT cascades in vivo. Dual luciferase reporter assay and RNA pull down assay were conducted to evaluate the interactions of lncRNA H19, p53 and TNFAIP8. Results The expression of lncRNA H19 and TNFAIP8 was up-regulated in breast cancer tissues and cell lines, especially in triple-negative breast cancer (TNBC). Functionally, knockdown of lncRNA H19 or TNFAIP8 coused the capacities of cell proliferation, migration, and invasion were suppressed, and cell cycle arrest was induced, as well as that the EMT markers were expressed abnormal. Mechanistically, lncRNA H19 antagonized p53 and increased expression of its target gene TNFAIP8 to promote EMT process. Furthermore, silencing of lncRNA H19 or TNFAIP8 also could inhibit tumorigenesis and lymph node metastases of MDA-MB-231 cells in xenograft nude mouse models. Conclusions Our findings provide insight into a novel mechanism of lncRNA H19 in tumorigenesis and metastases of breast cancer and demonstrate H19/p53/TNFAIP8 axis as a promising therapeutic target for breast cancer, especially for TNBC.

The Liver Transplant Risk Score (LTRS) is a simple clinical instrument developed to predict post liver transplant outcomes based on patient characteristics measured at the time of listing. The LTRS was developed using data of adult patients transplanted in the United States. In this study, we validated the performance of the LTRS in a cohort of patients transplanted in Europe.

Background Endomyocardial biopsy (EMB) is costly and discomforting yet remains a key component of surveillance after pediatric heart transplantation (HT). Donor-derived cell-free DNA (dd-cfDNA) has been histologically validated with high negative predictive value, offering an alternative to surveillance EMB (sEMB). Methods We implemented an alternative surveillance protocol using commercially available dd-cfDNA assays in place of sEMB after pediatric HT. Recipients ≧7 months post-HT with reassuring clinical assessment were referred for dd-cfDNA. When not elevated above the manufacturers' threshold, sEMB was deferred. Subsequent clinical status and results of follow-up EMB were analyzed. Results Over 17 months, 58 recipients [34% female, median age at HT 3.1 years (IQR 0.6-10.6)] had dd-cfDNA assessed per protocol. Median age was 14.8 years (8.4-18.3) and time from HT 6.0 years (2.2-11.2). Forty-seven (81%) had non-elevated dd-cfDNA and 11 (19%) were elevated. During a median of 8.7 months (4.2-15), all are alive without allograft loss/new dysfunction. Among those with non-elevated dd-cfDNA, 24 (51%) had subsequent sEMB at 12.1 months (6.9-12.9) with 23 showing no acute rejection (AR): grade 0R/pAMR0 (n = 16); 1R(1A)/pAMR0 (n = 7). One had AR (grade 2R(3A)/pAMR0) on follow-up sEMB after decreased immunosuppression following a diagnosis of PTLD. All 11 with elevated dd-cfDNA had reflex EMB at 19 days (12-32) with AR in 4: grade 1R(1B-2)/pAMR0 (n = 3); 1R(1B)/pAMR2 (n = 1). Conclusions dd-cfDNA assessment in place of selected, per-protocol EMB decreased surveillance EMB by 81% in our pediatric HT recipient cohort with no short-term adverse outcomes. Individual center approach to surveillance EMB will influence the utility of these findings.

Autoantibodies are detrimental to the survival of organ transplantation. We demonstrated that Angiotensin II Type I Receptor agonistic autoantibodies (AT1R-AA) were associated with poor outcomes after liver retransplantation. To examine the effect of other autoantibodies, we studied a retrospective cohort of 93 patients who received a second liver transplant. Pre-retransplant sera were tested with Luminex-based solid-phase assays. Among 33 tested autoantibodies, 15 were significantly higher in 48 patients who lost their regrafts than 45 patients whose regrafts were still functioning. Specifically, patients with autoantibodies to the C-terminal laminin-like globular domain of Perlecan (LG3) experienced significantly worse regraft survival (p = .002) than those with negative LG3 autoantibodies (LG3-A). In multivariate analysis, LG3-A (HR = 2.35 [1.11-4.98], p = .027) and AT1R-AA (HR = 2.09 [1.07-4.10], p = .032) remained significant predictors of regraft loss after adjusting for recipient age and sex. There were synergistic deleterious effects on regraft survival in patients who were double-positive for LG3-A and donor-specific antibody (DSA) (HR = 5.26 [2.15-12.88], p = .001), or LG3-A and AT1R-AA (HR = 3.23 [1.37-7.66], p = .008). All six double-positive patients lost their liver regrafts. In conclusion, LG3-A is associated with inferior long-term outcomes of a second liver transplant. Screening anti-HLA antibodies and autoantibodies such as LG3-A/AT1R-AA identifies patients with a higher risk for liver transplantation.

Recommender systems should be able to handle highly sparse training data that continues to change over time. Among the many solutions, Ant Colony Optimization, as a kind of optimization algorithm modeled on the actions of an ant colony, enjoys the favorable characteristic of being optimal, which has not been easily achieved by other kinds of algorithms. A recent work adopting genetic optimization proposes a collaborative filtering scheme: Ant Collaborative Filtering (ACF), which models the pheromone of ants for a recommender system in two ways: (1) use the pheromone exchange to model the ratings given by users with respect to items, (2) use the evaporation of existing pheromone to model the evolution of users&rsquo, preference change over time. This mechanism helps to identify the users and the items most related, even in the case of sparsity, and can capture the drift of user preferences over time. However, it reveals that many users share the same preference over items, which means it is not necessary to initialize each user with a unique type of pheromone, as was done with the ACF. Regarding the sparsity problem, this work takes one step further to improve the Ant Collaborative Filtering&rsquo, s performance by adding a clustering step in the initialization phase to reduce the dimension of the rate matrix, which leads to the results that K&lt, &lt, users, where K is the number of clusters, which stands for the maximum number of types of pheromone carried by all users. We call this revised version the Improved Ant Collaborative Filtering (IACF). Experiments are conducted on larger datasets, compared with the previous work, based on three typical recommender systems: (1) movie recommendations, (2) music recommendations, and (3) book recommendations. For movie recommendation, a larger dataset, MoviesLens 10M, was used, instead of MoviesLens 1M. For book recommendation and music recommendation, we used a new dataset that has a much larger size of samples from Douban and NetEase. The results illustrate that our IACF algorithm can better deal with practical recommendation scenarios that handle sparse dataset.

Human leukocyte antigen (HLA) class I presentation pathway plays a central role in natural killer (NK) cell and cytotoxic T-cell activities against BK polyomavirus (BKPyV) DNAemia. We determined the risk of sustained BKPyV DNAemia in 175 consecutive renal transplant recipients considering the simultaneous effect of donor/recipient HLA class I antigens and pre- or post-transplant variables. Median (IQR) age was 53 (44-64) years, and 37% of patients were female. 40 patients (22.9%) developed sustained BKPyV DNAemia [median (IQR) viral load: 9740 (4350-17 125) copies/ml]. In the Cox proportional hazard analysis, HLA-A1 (HR: 3.06, 95% CI: 1.51-6.17) and HLA-B35-Cw4 (HR: 4.63, 95% CI: 2.12-10.14) significantly increased the risk of sustained BKPyV DNAemia, while 2 HLA-C mismatches provided a marginally protective effect (HR: 0.32, 95% CI: 0.10-0.98). HLA-Cw4 is a ligand for NK cell inhibitory receptor, and HLA-B35 is in strong linkage disequilibrium with the HLA-Cw4 allele. The association between HLA-B35-Cw4 expression and sustained BKPyV DNAemia supports the important role of cytotoxic T cells and NK cells that would normally control BKPyV activation through engagement with immunoglobulin-like killer receptors (KIRs). Further studies are required to investigate the effect of HLA-C alleles along with NK cell activity against BKPyV DNAemia.

Background/Aims: Radiation therapy is an important and effective modality for the treatment of non-small cell lung cancer (NSCLC). MicroRNAs (miRNAs) are crucial post-transcriptional regulators that are involved in numerous important biologic processes. However, their potential involvement in radiation sensitivity remains unknown. Materials: We performed integrated analysis of miRNA expression in NSCLC using The Cancer Genome Atlas datasets. miR-99a was found to be significantly upregulated in cancer tissue and regulated cell survival. Cell culture was used to assess the role of miR-99a in radiation sensitivity. We then used flow cytometry to examine the effects of miR-99a on the cell cycle and apoptosis in cells exposed to radiation. To identify gene targets of miR-99a, a bioinformatics approach was adopted, and the findings of this analysis were verified using luciferase reporter assays. Finally, an in vivo study was conducted to examine the effect of miR-99a on tumor volume in an NSCLC mouse model undergoing radiation therapy. Results: miR-99a was significantly upregulated in radiation-sensitive A549 cells compared with radiation-resistant A549 cells. miR-99a overexpression was shown to enhance radiosensitivity, while inhibition of miR-99a resulted in radioresistance of NSCLC cell lines in vitro and in vivo. In addition, by bioinformatics software analysis and luciferase assays, mammalian target of rapamycin (mTOR) was identified as a direct target of miR-99a. Furthermore, AZD2014, an inhibitor of mTOR, enhanced radiosensitivity and apoptosis in NSCLC cell lines, while mTOR overexpression resulted in radioresistance and cell survival from miR-99a-induced cell apoptosis. Moreover, miR-99a overexpression further increased the efficacy of radiation therapy in an NSCLC xenograft mouse model, and miR-99a and mTOR expression was significantly inversely correlated. Conclusions: Altogether, these data suggested miR-99a functions as a tumor suppressor that has a critical role in regulating radiosensitivity of NSCLC by targeting the mTOR signaling pathway.

With the increasing amount of high-resolution remote sensing images, it becomes more and more urgent to retrieve remote sensing images from large archives efficiently. The existing methods are mainly based on shallow features to retrieve images, while shallow features are easily affected by artificial intervention. Recently, convolutional neural networks (CNNs) are capable of learning feature representations automatically, and CNNs pre-trained on large-scale datasets are generic. This paper exploits representations from pre-trained CNNs for high-resolution remote sensing image retrieval. CNN representations from AlexNet, VGGM, VGG16, and GoogLeNet are first transferred for high-resolution remote sensing images, and then CNN features are extracted via two approaches. One is extracting the outputs of high-level layers directly and the other is aggregating the outputs of mid-level layers by means of average pooling with different pooling regions. Given the generalization and high dimensionality of the CNN features, feature combination and feature compression are also adopted to improve the feature representation. Experimental results demonstrate that aggregated features with pooling region smaller than the feature map size perform excellently, especially for VGG16 and GoogLeNet. Shallow feature makes a great contribution to enhance the retrieval precision when combined with CNN features, and compressed features reduce redundancy effectively. Compared with the state-of-the-art methods, the proposed feature extraction methods are very simple, and the features are able to improve retrieval performance significantly.

The effect of postmastectomy radiotherapy (PMRT) on T1-2 breast cancer patients with 1-3 positive axillary lymph nodes is controversial up to now. The purpose of this study was to evaluate the impact of postmastectomy radiotherapy for these patients. The prognostic factor effecting locoregional free-survival (LRFS) was also analyzed. In the retrospective clinical data of 1674 eligible patients, survival analysis was performed using the method of Kaplan-Meier and the log-rank test. Cox regression analysis was applied to identify the significant prognostic factors. We found PMRT increased 5-year LRFS (p=0.003), but could not improve 5-year disease-free survival or overall survival statistically. For patients without PMRT, multivariate analysis revealed that age, lymph node ratio and molecule subtype were risk factors effecting LRFS. To further analyze the role of PMRT, we grouped all the patients into low risk group (0 or 1 risk factor) and high risk group (2 or 3 risk factors) depending on these risk factors. We found that in low-risk group, PMRT increased only 5-year LRFS (p=0.012). However, in high-risk group, PMRT increased both 5-year LRFS (p=0.005) and 5-year disease-free survival (p=0.033), but could not improve 5-year overall survival statistically. Thus, these data provide the evidence that PMRT could improve LRFS for T1-2 breast cancer patients with 1-3 positive axillary lymph nodes. Additionally, PMRT could improve LRFS and disease-free survival for high risk patients. Age, lymph node ratio and molecule subtype were high risk factors effecting LRFS in our study.

Summary Tumor necrosis factor α –induced protein 8 (TNFAIP8), a transcription factor nuclear factor κ B–inducible, antiapoptotic and oncogenic molecule, is associated with prognosis of several human malignancies. However, the relationship between TNFAIP8 and the prognosis of the invasive ductal carcinoma (IDC) of the breast remains unclear. TNFAIP8 expression was evaluated using real-time polymerase chain reaction (PCR) and Western blot analysis in 20 fresh IDC tissues and immunohistochemical analysis in 351 paraffin-embedded IDC tissues. Real-time PCR and Western blot analysis demonstrated that both TNFAIP8 messenger RNA and protein were up-regulated in IDC tissues compared with the paired adjacent noncancerous tissues. Immunohistochemistry revealed that TNFAIP8 expression was significantly correlated with some clinicopathological factors, including axillary lymph node metastasis ( P =.001), advanced TNM stage ( P .001), high histologic grade ( P .001), molecular subtype ( P .001), and postoperative recurrence ( P .001). Univariate and multivariate logistic regression analyses demonstrated that TNFAIP8 overexpression was strongly associated with axillary lymph node metastasis (odds ratio, 1.818; 95% confidence interval, 1.167-2.832; P =.008). Moreover, Kaplan-Meier analysis indicated that IDC patients with high TNFAIP8 expression had a shorter survival time than did those with low TNFAIP8 expression, and multivariate analysis indicated that TNFAIP8 was an independent prognostic factor for overall survival and disease-free survival in IDC ( P =.041 and P =.020, respectively). Therefore, TNFAIP8 overexpression may contribute to tumor progression, and it may be a novel prognostic biomarker for the patients with IDC.

Angiotensin II type I receptor (AT1R) agonistic autoantibodies (AT1R-AA) are detrimental to kidney transplantation. Early studies suggested a similar negative effect in primary liver transplantation. Here, we studied AT1R-AA in a retrospective cohort of 94 patients who received a second liver transplant to determine their prevalence and effects. The concentrations of preformed AT1R-AA before transplantation were higher (P = .019) in the 48 patients who lost their liver grafts than in the 46 patients whose grafts survived. About half (48/94, 51.1%) of the patients were positive for AT1R-AA17 U/mL before the second liver transplantation. In 22 (23.4%) patients, strong positive AT1R-AA (defined as40 U/mL) were detected, of whom 16 (72.7%) patients lost their grafts. Based on Kaplan-Meier analysis, patients with strong positive AT1R-AA had significantly worse graft survival than those with AT1R-AA40 U/mL (P = .035). In multivariate Cox models that included confounders such as sex and age, either AT1R-AA40 U/mL (HR = 1.999 [1.085-3.682], P = .026) or increased concentrations of AT1R-AA (HR = 1.003 [1.001-1.006] per incremental U/mL, P = .019) were significantly associated with elevated risk for graft loss. In conclusion, our data indicate that there is a high prevalence of AT1R-AA in candidates for second liver transplantation and that their presence is associated with inferior long-term outcomes of the second graft.

Human leukocyte antigen (HLA) compatibility is very important for successful transplantation of solid organs. In this paper, we focused on the humoral arm of immunity in the clinical setting of organ transplantation: how HLA antibodies develop, how they can be detected, and what they can do to injure organ transplants. Specifically, we explore the technical perspectives of detecting donor-specific antibodies (DSA) in HLA laboratories, and use real-life clinical cases to explain the principles. Currently there are many tools in our HLA antibody detection toolbox: conventional cytotoxicity cross match, flow cross match, and solid phase assays using beads conjugated with single or multiple HLA antigens. Single antigen bead (SAB) assay is the most sensitive tool available for detecting HLA antibodies and assessing the immunological risk for organ transplant. However, there are intrinsic limitations to solid-phase assays and they are prone to both false negativity and importantly, false positivity. Denatured antigens on single antigen beads might be the most prominent source of false positive reactivity, and may have been underestimated by many HLA experts. No single assay is perfect and therefore multiple methods, including the less sensitive assays, should be employed to determine the clinical relevance of detected HLA antibodies. Thoughtful process, including knowledge of HLA systems, cross reactivity, epitopes, and the patient’s clinical history should be employed to correctly interpret data. The clinical team should work closely with HLA laboratories to ensure accurate interpretation of information and optimal management of patients before and after organ transplantation.

Background: Despite reports that associate donor specific antibody (DSA) with rejection after liver transplantation, grafts are still allocated according to blood group (ABO) but not human leukocyte antigen (HLA) compatibility, possibly due to the absence of an easily discernible clinical association between adverse recipient outcome and DSA. Re-transplantation provides a test environment where the presence of preformed DSA is prevalent and its effect on outcome should be apparent. Methods: All patients undergoing a second liver transplantation with available pre-operative serum were included with the exception of ABO incompatible or multiple organ transplants. Banked sera were tested for anti-HLA antibodies with Luminex-based solid phase assays. Anti-HLA antibodies to the second donor (D2SA) were determined using antibodies specificity and HLA typing of 2nd liver donor. Results: Preformed HLA antibodies directed to second liver transplantation (D2SA) were found in 31 (39%) of the 79 patients that were included in the study. Primary and re-transplantation characteristics were similar in both subgroups except first graft survival which was significantly shorter in recipients who are negative for D2SA. Mean survival of the second graft was similar in D2SA+ and D2SA− cohorts [8.55 (range, 0.01–24.74) vs . 7.56 (range, 0–23.53) years respectively, P=0.574]. Mean patient survival after 2nd liver transplantation was similar in D2SA+ and D2SA− cohorts [9.11 (range, 0.01–24.74) vs . 8.10 (range, 0–23.53) years respectively, P=0.504]. Subgroup univariate analysis demonstrated no detrimental effect of class, locus, or strength of D2SA on survival of the second liver transplant. In multivariate cox regression model, neither class I D2DSA (HR =1.101, P=0.92) nor class II D2SA (HR =1.74, P=0.359) were significant risks of graft failure. Conclusions: Presence of D2SA was not found to be associated with inferior outcomes in this retrospective cohort study of liver re-transplantation suggesting that changes to the allocation system are not required.

Despite implementation of virtual crossmatches, flow cytometry crossmatches (FCXM) are still used by many transplant centers to determine immunological risk before kidney transplantation. To determine if common profiles of patients prone to false positive FCXM exist, we examined the demographics and native diseases of kidney patients tested with autologous FCXM (n = 480). Improvements to FCXM and cell isolation methods significantly reduced the positive rate from 15.1% to 5.3%. Patients with native diseases considered ‘immunological’ (vasculitis, lupus, IgA nephropathy) had more positive autologous FCXM (OR = 3.36, p = 0.003) vs. patients with all other diseases. Patients who were tested using our updated method (n = 321) still showed that these immunological diseases were a significant predictor for positive autologous FCXM (OR = 4.79, p = 0.006). Interestingly, patients on peritoneal dialysis (PD) also had significantly more positive autologous FCXM than patients on hemodialysis or waiting for pre-emptive kidney transplants (OR = 3.27, p = 0.02). These findings were confirmed in patients who had false positive allogeneic FCXM. Twenty of 24 (83.3%) patients with false positive allogeneic FCXM tested with updated method either had immunological diseases originally or were on PD. Our findings are helpful when interpreting an unexpected positive FCXM, especially for transplantation from deceased donors.

Introduction Antibody-mediated rejection (AMR) is an increasingly recognized cause of lung transplant allograft failure. Treatment of AMR involves combination therapy that is often poorly tolerated or unsuccessful with regimens involving the combination of intravenous immunoglobulins, plasmapheresis, and proteasome inhibitors, complement, or CD20 therapy. The development of DSAs alone, specifically complement activating C1q-binding DSAs, has been identified as an independent risk factor for solid organ transplant allograft dysfunction. Previous studies have examined the efficacy of combination therapy involving the anti-CD20 antibody rituximab to preemptively eliminate DSAs. Here we report on a case of successful eradication of de novo C1q-binding DSAs in a lung transplant recipient using rituximab. Case Report Our patient underwent double-lung transplantation for idiopathic pulmonary fibrosis in August 2019. His post-transplant course was notable for multiple episodes of acute cellular rejection (ACR) requiring burst-dose corticosteroids. DSA titers were monitored every three months post-transplant. At six months, de novo DSAs were detected (DQ7 with DQA1*05) at a mean fluorescence intensity (MFI) of 3623 (low-moderate) (Figure 1). These subsequently rose to an MFI of 11281 (strong) with development of C1q positivity that peaked at an MFI of 15038. Given his frequent episodes of ACR, the strong C1q positivity, and concerns regarding his ability to tolerate therapy for potential future AMR, he was preemptively treated with a single infusion of rituximab (375 mg per meter-squared body surface area). Follow-up DSA titers demonstrated near-total elimination of the C1q component (MFI 403). The patient tolerated the treatment without issue and continues to demonstrate improvement in his allograft function. Summary Rituximab monotherapy can be considered preemptively in patients with C1q-binding DSAs that may not tolerate traditional treatment of antibody-mediated rejection.

Purpose Detection of donor-derived cell-free DNA (dd-cfDNA) in circulating blood of posttransplant recipients is a clinically informative tool to predict acute allograft injury and rejection, although its reliability and feasibility have not been extensively studied in lung transplantation. Here, we present our preliminary data of dd-cfDNA in lung transplant patients during the early posttransplant phase. Methods Eight subjects were included in this study among patients received double lung transplants at our institute between Jun.-Sept. 2020. Re-transplantation cases were excluded per the study protocol. Blood samples were collected at various times from these consented patients before and after transplantation. Dd-cfDNA in obtained blood samples were determined using AlloSure dd-cfDNA test kits (CareDx, Inc). Results Of the 8 subjects, none developed severe primary graft dysfunction and two patients presented acute cellular rejection (ACR) after lung transplantation. All subjects exhibited increased dd-cfDNA immediately after transplantation with a peak percentage of dd-cfDNA (range 12-27%) within 48h posttransplant. (Figure 1). Over 2 weeks, dd-cfDNA percentage declined with significant variation reaching a similar level of Conclusion To our knowledge, this is the first report of acute time-dependent changes of dd-cfDNA in patients immediately after lung transplantation. Our preliminary data suggest that recipients’ systemic condition may mask the appearance of dd-cfDNA. Additional information regarding circulating whole cfDNA in patients may assist in more accurate detection of dd-cfDNA in the acute phase.

Decorin, chiefly synthesized by tumor stroma, is known as a tumor suppressor. However, the clinical and prognostic significance in lung cancer remained unclear. Here, decorin and Ki67 expression was detected by immunohistochemistry (IHC) in I–IIIA non-small cell lung cancer (NSCLC) tissues (n = 264) in comparison to adjacent normal tissues (n = 40). The relationship between the expression of decorin and clinical characteristics, as well as Ki67 index and prognosis, was analyzed. Decorin expression was decreased in both the stroma (P

Purpose Allosensitization in heart transplant (HT) candidates is associated with longer transplant wait times and post-transplant complications. We summarize our experience with desensitization using carfilzomib (CFZ), an irreversible proteasome inhibitor that causes plasma cell apoptosis. Methods One cycle of desensitization consisted of plasmapheresis and CFZ 20mg/m2 on days 1, 2, 8, 9, 15, and 16 with intravenous immune globulin 2g/kg after CFZ on day 16. Patients underwent repeat cycles as indicated. We summarize treatment impact by depicting calculated PRA (cPRA) for neat combined Class I and II IgG and C1q pre- and post-treatment using a cutoff for cPRA entry of ≥ 4000 and 500 MFI, respectively. We fit a linear mixed model to evaluate treatment effect. Results From June 2013 to October 2019, 9 patients underwent 20 cycles of CFZ with 6 requiring multiple cycles. Each cycle of CFZ resulted in an average cPRA decrease of 33% (95% CI: 16-50) for combined IgG and 36% (95% CI: 17-55) for C1q. An example is shown in Figure 1. From treatment start to finish, mean cPRA fell from 76% to 35% (p=0.005) for combined IgG and 56% to 4% (p=0.017) for C1q (Table 1). Six of 9 patients have been transplanted with 5 of the transplanted hearts crossing historical antibodies. HT survival at 1 year was 100%. Patient 6 has not been transplanted due to obesity. Patients 3 and 4 only received 4 doses of CFZ due to acute kidney injury, which resolved. Neutropenia, thrombocytopenia, neuropathy, and hepatotoxicity did not occur. Conclusion CFZ, as part of a desensitization regimen, appears to be safe and effective in sensitized patients awaiting HT. The number of cycles may depend on the HLA antibody titer prior to treatment.

The aim of our study is to construct the competing endogenous RNA (ceRNA) network of head and neck squamous cell carcinoma (HNSCC) and identify key long noncoding RNAs (lncRNAs) to predict prognosis. The genes whose expression were differentially in HNSCC and normal tissues were explored by the Cancer Genome Atlas database. The ceRNA network was constructed by the Cytoscape software. The lncRNAs which could estimate the overall survival were explored from Cox proportional hazards regression. There are 1997, 589, and 82 mRNAs, lncRNAs, and miRNAs whose expression were statistically significant different, respectively. Then, the network between miRNA and mRNA or miRNA and lncRNA was constructed by miRcode, miRDB, TargetScan, and miRanda. Five mRNAs, 10 lncRNAs, and 3 miRNAs were associated with overall survival. Then, 11-lncRNAs were found to be prognostic factors. Therefore, our research analyzed the potential signature of novel 11-lncRNA as candidate prognostic biomarker from the ceRNA network for patients with HNSCC.

In the last decades, Content Based Image Retrieval and image classification have become popular, and among them Region-based Image Retrieval is quite active. More and more descriptors and retrieval methods have been proposed and investigated in order to improve the retrieval performance. This paper proposed a feature fusion deep learning method. The features including colors, texture and shape, which are extracted from both the entire image and regions. The features are then trained using diverse deep learning methods. The conducted deep learning methods include Sparse Auto-encoders, Denoising Autoencoding, Deep Belief Nets, Drop Out Neural Networks, and Deep Boltzmann Machine. The method is evaluated through extensive experiments on Corel 10K datasets. Experimental results demonstrate that the introduced methods are comparable with the state-of-arts in this image retrieval application.

Wavelet packet multicarrier system gains widespread concern because of its better resistance performance to Inter-Symbol Interference (ISI) and Inter-Carrier Interference (ICI), as well as the higher spectrum efficiency. However, multicarrier system has a high Peak to Average Power Ratio (PAPR), which will lead to many problems such as lower system performance. In order to solve the problem, a kind of PAPR reduction method based on pruning Wavelet Packet Modulation (WPM) and Partial Transmit Sequences (PTS) technology is proposed in this paper, through proper pruning of the full-tree structure of wavelet packet modulation in the PTS technology to reduce the number of nodes in the system, and finally improve the reduction effect of PAPR. Simulation results show that when Complementary Cumulative Distribution Function (CCDF) is 10−3, PTS based on pruning WPM compared with PTS technique and pruning technique has improved about 1 dB and 1.5 dB, which will not affect the system’s Bit Error Rate (BER) performance in the wavelet packet multicarrier system.

Increased CD8(+) T-cell precursor frequency (PF) precludes the requirement of CD4(+) helper T (Th) cells for primary CD8(+) cytotoxic T-lymphocyte (CTL) responses. However, the key questions of whether unhelped CTLs generated at higher PF are functional effectors, and whether unhelped CTLs can differentiate into functional memory cells at higher PF are unclear. In this study, ovalbumin (OVA) -pulsed dendritic cells (DC(OVA)) derived from C57BL/6, CD40 knockout (CD40(-/-)) or CD40 ligand knockout (CD40L(-/-)) mice were used to immunize C57BL/6, Ia(b-/-), CD40(-/-) or CD40L(-/-) mice, whose PF was previously increased with transfer of 1 × 10(6) CD8(+) T cells derived from OVA-specific T-cell receptor (TCR) transgenic OTI, OTI(CD40(-/-)) or OTI(CD40L(-/-)) mice. All the immunized mice were then assessed for effector and memory CTL responses. Following DC immunization, relatively comparable CTL priming occurred without CD4(+) T-cell help and Th-provided CD40/CD40L signalling. In addition, the unhelped CTLs were functional effectors capable of inducing therapeutic immunity against established OVA-expressing tumours. In contrast, the functional memory development of CTLs was severely impaired in the absence of CD4(+) T-cell help and CD40/CD40L signalling. Finally, unhelped memory CTLs failed to protect mice against lethal tumour challenge. Taken together, these results demonstrate that CD4(+) T-cell help at higher PF, is not required for effector CTL priming, but is required for functional memory CTL development against cancer. Our data may impact the development of novel preventive and therapeutic approaches in cancer patients with compromised CD4(+) T-cell functions.