An Efficient Probe-Based Quantitative PCR Assay Targeting Human-Specific DNA in ST6GALNAC3 for the Quantification of Human Cells in Preclinical Animal Models.

Precise quantification of human cells in preclinical animal models by a sensitive and specific approach is warranted. The probe-based quantitative PCR (qPCR) assay as a sensitive and swift approach is suitable for the quantification of human cells by targeting human-specific DNA sequences. In this study, we developed an efficient qPCR assay targeting human-specific DNA in ST6GALNAC3 (termed ST6GAL-qPCR) for the quantification of human cells in preclinical animal models. ST6GAL-qPCR probe was synthesized with FAM and non-fluorescent quencher-minor groove binder conjugated to the 5' and 3' end of the probe, respectively. Genomic DNA from human, rhesus monkeys, cynomolgus monkeys, New Zealand White rabbits, SD rats, C57BL/6, and BALB/c mice were utilized for analyzing the specificity and sensitivity of the ST6GAL-qPCR assay. The ST6GAL-qPCR assay targeted human-specific DNA was cloned to pUCM-T vector and released by EcoR I/Hind III digestion for generating a calibration curve. Cell mixing experiment was performed to validate the ST6GAL-qPCR assay by analysis of 0.1%, 0.01%, and 0.001% of human leukocytes mixed with murine thymocytes. The ST6GAL-qPCR assay detected human DNA rather than DNA from the tested animal species. The amplification efficiency of the ST6GAL-qPCR assay was 93% and the linearity of calibration curve was R2 = 0.999. The ST6GAL-qPCR assay detected as low as 5 copies of human-specific DNA and is efficient to specially amplify as low as 30-pg human DNA in the presence of 1 µg of DNA from the tested species, respectively. The ST6GAL-qPCR assay was able to quantify as low as 0.01% of human leukocytes within murine thymocytes. This ST6GAL-qPCR assay can be used as an efficient approach for the quantification of human cells in preclinical animal models.

A nomogram for predicting breast cancer specific survival in elderly patients with breast cancer: a SEER population-based analysis.

BACKGROUND: The number of elderly patients diagnosed with breast cancer is increasing worldwide. However, treatment decisions for these patients are highly variable. Although researchers have identified the effects of surgery, radiotherapy, endocrine therapy, and chemotherapy in elderly patients with breast cancer, clinicians still struggle to make appropriate decisions for these patients. METHODS: We identified 75,525 female breast cancer patients aged ≥ 70 years in the Surveillance, Epidemiology, and End Results (SEER) database treated between January 1, 2010, and December 31, 2016. The patients were further divided into training and testing cohorts. The cumulative occurrence of breast cancer-specific deaths (BCSDs) and other cause-specific deaths (OCSD) was calculated using the cumulative incidence function. In the univariate analysis, risk factors were screened using the Fine-Gray model. In the multivariate analysis for competing risks, the sub-distribution hazard ratio with a 95% confidence interval for each independent predictor associated with BCSD was calculated for the construction of nomograms. Based on the above analyses, a competing risk nomogram was constructed to predict the probability of BCSD in the 1st, 3rd, and 5th years after treatment. During validation, the concordance index (C-index) was selected to quantify the predictive ability of the competing risk model. RESULTS: A total of 33,118 patients were included in this study, with 24,838 in the training group and 8,280 in the testing group. Age, race, marital status, cancer grade, tumor stage, node stage, estrogen receptor status, progesterone receptor status, human epidermal growth factor receptor--2 status, and treatment including surgery, radiation, and chemotherapy were used to establish a nomogram. The C-index of 0.852 (0.842-0.862) in the training cohort and 0.876 (0.868-0.892) in the testing cohort indicated satisfactory discriminative ability of the nomogram. Calibration plots showed favorable consistency between the nomogram predictions and actual observations in both the training and validation cohorts. CONCLUSIONS: Our study identified independent predictors of BCSD in elderly patients with breast cancer. A prognostic nomogram was developed and validated to aid clinical decision-making.

Diagnostic value of core needle biopsy for determining HER2 status in breast cancer, especially in the HER2-low population.

PURPOSE: The status of human epidermal growth factor receptor 2 (HER2) is important for treatment decision-making of breast cancer and was commonly determined by core needle biopsy (CNB). The concordance of CNB with surgical excision biopsy (SEB) has been verified, but remain unclear according to the newly developed classification of HER2 status. Our study aimed to re-evaluate the diagnostic value of CNB for determining HER2 status in breast cancer, especially in the HER2-low population. METHODS: Eligible breast cancer patients in West China Hospital between January 1, 2007 and December 31, 2021 were enrolled consecutively and data were extracted from the Hospital Information System. The agreement of HER2 status between CNB and SEB was calculated by concordance rate and κ statistics, as well as the sensitivity, specificity, positive, and negative predictive values (PPV & NPV). Logistic models were used to explore potential factors associated with the discordance between both tests. RESULTS: Of 1829 eligible patients, 1097 (60.0%) and 1358 (74.2%) were consistent between CNB and SEB by pathological and clinical classifications, respectively, with κ value being 0.46 (0.43-0.49) and 0.57 (0.53-0.60). The sensitivity (50.9%-52.7%) and PPV (50.5%-55.2%) of CNB were especially low among IHC 1+ and 2+/ISH - subgroups by pathological classifications; however, it showed the highest sensitivity (77.5%) and the lowest specificity (73.9%) in HER2-low population by clinical classifications. Advanced N stages might be a stable indicator for the discordance between both tests. CONCLUSION: The diagnostic value of CNB was limited for determining HER2 status in breast cancer, especially in HER2-low population.

Effect of adjuvant chemotherapy on the survival outcomes of elderly breast cancer: A retrospective cohort study based on SEER database.

BACKGROUND: Currently, the proportion of standard chemotherapy for elderly patients is much lower than that for young patients, with little evidence from clinical trials supporting the use of chemotherapy for elderly patients. The effectiveness of chemotherapy for the elderly suffering from breast cancer remains to be further verified. METHODS: A total of 75,525 female breast cancer patients aged 70 years or older were hereby identified, all from the Surveillance, Epidemiology, and End Results (SEER) database from January 1, 2010 to December 31, 2016. Kaplan-Meier analysis and multivariable Cox proportional model were performed to evaluate the effectiveness of chemotherapy on overall survival (OS) and breast cancer-specific survival (BCSS). Propensity score matching (PSM) (PSM ratio: 1:1, caliper: 0.2 standard deviation of propensity score) was applied to construct balanced cohorts with or without chemotherapy based on demographic and pathophysiological characteristics. RESULTS: A total of 33,177 eligible patients were included, with 5273 (15.89%) receiving chemotherapy. Through PSM, 8360 patients were successfully matched, and balances between groups were almost reached. In the matched data set, multivariable Cox analysis reveals that chemotherapy was associated with a 36% and 21% risk reduction on OS (HR = 0.64, 95% CI 0.58 to 0.71) and BCSS (HR = 0.79, 95% CI 0.69 to 0.91), respectively. Furthermore, subgroups with more adjacent lymph nodes involved by tumor, or nonluminal A, were inclined to benefit more from chemotherapy. Moreover, chemotherapy did not increase the chances of dying from heart disease. CONCLUSIONS: The present study provided evidence that chemotherapy may improve the prognosis of elderly breast cancer, especially for those subpopulations that benefit more from chemotherapy treatment.

COVID-19 Vaccination Status and Hesitancy among Breast Cancer Patients after Two Years of Pandemic: A Cross-Sectional Survey.

Background: Patients with cancer show greater susceptibility and vulnerability to severe acute respiratory syndrome coronavirus 2 infection. However, data on the vaccination status among patients with breast cancer and any structured analysis of the factors influencing patients' decisions regarding vaccines are lacking. Methods: This cross-sectional study on patients with breast cancer in China was conducted from 1 June 2022, to 17 June 2022. Every participant completed an online questionnaire about their vaccination status and any adverse reactions, and a scale based on the Health Belief Model (HBM) to assess the vaccination status of respondents and their willingness to receive following doses or boosters. Results: Among the 1132 participants, 55.2% had received a COVID-19 vaccine. The incidence of adverse events per dose was around 40%. Vaccine hesitancy of 61.9% was observed among patients who had not fully received three doses of vaccine or boosters. The only variable found to be associated with vaccine hesitancy was time since diagnosis (p < 0.05). In the HBM scale, vaccine hesitancy was closely related to a low level of perceived susceptibility, a low level of perceived benefit, a high level of perceived barriers and a low level of agreement with doctors' advice. Conclusions: For patients with breast cancer, perceived susceptibility, benefits and barriers should be prioritized, and the advice from authoritative doctors is a vital cue to action.

Therapeutic progress and challenges for triple negative breast cancer: targeted therapy and immunotherapy.

Triple negative breast cancer (TNBC) is a subtype of breast cancer, with estrogen receptor, human epidermal growth factor receptor 2 and progesterone receptor negative. TNBC is characterized by high heterogeneity, high rates of metastasis, poor prognosis, and lack of therapeutic targets. Now the treatment of TNBC is still based on surgery and chemotherapy, which is effective only in initial stage but almost useless in advanced stage. And due to the lack of hormone target, hormonal therapies have little beneficial effects. In recent years, signaling pathways and receptor-specific targets have been reported to be effective in TNBC patients under specific clinical conditions. Now targeted therapies have been approved for many other cancers and even other subtypes of breast cancer, but treatment options for TNBC are still limited. Most of TNBC patients showed no response, which may be related to the heterogeneity of TNBC, therefore more effective treatments and predictive biomarkers are needed. In the present review, we summarize potential treatment opinions for TNBC based on the dysregulated receptors and signaling pathways, which play a significant role in multiple stages of TNBC development. We also focus on the application of immunotherapy in TNBC, and summarize the preclinical and clinical trials of therapy for patients with TNBC. We hope to accelerate the research and development of new drugs for TNBC by understanding the relevant mechanisms, and to improve survival.

Safety and efficacy profile of mogamulizumab (Poteligeo) in the treatment of cancers: an update evidence from 14 studies.

BACKGROUND: CC chemokine receptor 4 (CCR4), the receptor for CCL22 and CCL17, is expressed on the surface of effector Tregs that have the highest suppressive effects on antitumor immune response. CCR4 is also widely expressed on the surface of tumor cells from patients with adult T-cell leukemia/lymphoma (ATL), peripheral T-cell lymphoma (PTCL) and cutaneous T-cell lymphoma (CTCL). Mogamulizumab is a humanized, IgG1 kappa monoclonal antibody that is directed against CCR4. By reducing the number of CCR4-positive Tregs and tumor cells, the mogamulizumab can reduce tumor burden and boost antitumor immunity to achieve antitumor effects. METHODS: We examined the PubMed and ClinicalTrials.gov until 1 February 2020. Considering variability in different studies, we selected the adverse events (AEs), overall survival (OS), progression-free survival (PFS), objective responses rate (ORR) and Hazard Ratio (HR) for PFS to evaluate the safety and efficacy profile of mogamulizumab. RESULTS: When patients were treated with mogamulizumab monotherapy, the most common all-grade AEs were lymphopenia, infusion reaction, fever, rash and chills while the most common grade ≥ 3 AEs were lymphopenia, neutropenia and rash. When patients were treated with combined therapy of mogamulizumab and other drugs, the most common all-grade AEs were neutropenia, anaemia, lymphopenia and gastrointestinal disorder, while the most common grade ≥ 3 AEs was lymphopenia. For patients treated with mogamulizumab monotherapy, the pooled ORR and mean PFS were 0.430 (95% CI: 0.393-0.469) and 1.060 months (95% CI: 1.043-1.077), respectively. For patients treated with combined therapy of mogamulizumab and other drugs, the pooled ORR was 0.203 (95% CI: 0.022-0.746) while the pooled PFS and OS were 2.093 months (95% CI: 1.602-2.584) and 6.591 months (95% CI: 6.014-7.167), respectively. CONCLUSIONS: Based on present evidence, we believed that mogamulizumab had clinically meaningful antitumor activity with acceptable toxicity which is a novel therapy in treating patients with cancers.

Distinct Characteristics of COVID-19 Infection in Children.

SARS-CoV-2, a member of the family coronaviridae, has triggered a lethal pandemic termed coronavirus disease 2019 (COVID-19). Pediatric patients, mainly from families with a cluster of infection or a history of exposure to epidemic areas, get infected via direct contacts or air-borne droplets. Children (aged below 18 years) are susceptible to COVID-19, with an average incubation period of about 6.5 days. Most cases present asymptomatic or common cold symptoms such as fever, cough, and myalgia or fatigue, which is milder than adult patients. Besides, most abnormal laboratory and radiologic findings in children with COVID-19 are non-specific. Since no specific chemotherapeutic agents have been approved for children, timely preventive methods could effectively forestall the transmission of SARS-CoV-2. To date, mostly studied cases have been adults with COVID-19, whereas data on pediatrics patients remain poorly defined. We herein conducted a literature review for papers published in PubMed and medRxiv (preprints) between December 2019 and December 2020 that reported on pediatrics patients (aged below 18 years) with a confirmed COVID-19 diagnosis. In this review, we summarized and discussed the pathogenesis, epidemiology, and clinical management of COVID-19 in pediatrics patients to improve our understanding of this new disease in children.

Pattern of Time-to-Surgery in Patients With Breast Cancer at Different Stages of the COVID-19 Pandemic.

BACKGROUND: The management of cancer surgeries is under unprecedented challenges during the COVID-19 pandemic, and the breast cancer patients may face a time-delay in the treatment. This retrospective study aimed to present the pattern of time-to-surgery (TTS) and analyze the features of breast cancer patients under the different stages of the COVID-19 pandemic. METHODS: Patients who received surgeries for breast cancers at West China Hospital between February 15, 2020 and April 30, 2020 (the outbreak and post-peak stages), and between March 10, 2021 and May 25, 2021 (the normalization stage) were included. TTS was calculated as the time interval between the pathological diagnosis and surgical treatment of breast cancer patients. And the pandemic was divided into three stages based on the time when the patients were pathologically diagnosed and the severity of pandemic at that time point. TTS, demographic and clinicopathological features were collected from medical records. RESULTS: A total of 367 patients were included. As for demographic features, it demonstrated statistically significant differences in insurance type (p<0.001) and regular screening (p<0.001), as well as age (p=0.013) and menstrual status (p=0.004). As for clinicopathological features, axillary involvement (p=0.019) was a factor that differed among three stages. The overall TTS was 23.56 ± 21.39 days. TTS for patients who were diagnosed during the outbreak of COVID-19 were longer than those diagnosed during pandemic post-peak and normalization stage (p<0.001). Pandemic stage (p<0.001) and excision biopsy before surgery (OR, 6.459; 95% CI, 2.225-18.755; p=0.001) were markedly correlated with the TTS of patients. CONCLUSIONS: TTS of breast cancer patients significantly varied in different stages of the COVID-19 pandemic. And breast cancer patients' daily lives and disease treatments were affected by the pandemic in many aspects, such as health insurance access, physical screening and change of therapeutic schedules. As the time-delay may cause negative influences on patients' disease, we should minimize the occurrence of such time-delay. It is vital to come up with comprehensive measures to deal with unexpected situations in case the pandemic occurs.

Comparison of Diagnostic Performance of Five Different Ultrasound TI-RADS Classification Guidelines for Thyroid Nodules.

OBJECTIVES: We aimed to evaluate and compare the diagnostic performance of five ultrasound thyroid imaging reporting and data system (TI-RADS) classification guidelines for thyroid nodules through a review and meta-analysis. METHODS: We searched for relevant studies before February 2020 in PubMed. Then we pooled the sensitivity, specificity, likelihood ratios, diagnostic odds ratios, and area under the summary receiver operating characteristic curves. And the diagnostic odds ratios were used to compare the performance. RESULTS: We totally included 19 studies with 4,696 lesions in this research. The pooled sensitivity of American College of Radiology (ACR) guidelines, American Thyroid Association (ATA) guidelines, TI-RADS proposed by Kwak (Kwak TI-RADS), Korean Thyroid Association/Korean Society of Thyroid Radiology (KTA/KSThR) guidelines for malignancy risk and European Thyroid Association (ETA) guidelines is between 0.84 and 0.94. The pooled specificity is 0.68, 0.44, 0.62, 0.47, and 0.61, respectively. And the RDOR is 1.57 (ACR vs ATA), 1.37 (ACR vs ETA), 1.80 (ACR vs Kawk), 1.74 (ARC vs KTA). CONCLUSIONS: The results suggest that five classification guidelines are all effective methods for differential diagnosis of benign and malignant thyroid nodules and ACR guideline is a better choice.

68Ga-PSMA PET/MRI for the diagnosis of primary and biochemically recurrent prostate cancer: A meta-analysis.

PURPOSE: Our meta-analysis aimed to evaluate the diagnostic performance of 68Ga-labelled prostate-specific membrane antigen ligand positron emission tomography/magnetic resonance imaging (68Ga-PSMA PET/MRI) in patients with primary and biochemically recurrent prostate cancer (PCa). METHODS: We searched for relevant articles in PubMed, EMBASE, the Cochrane Library and Web of Science until September 12, 2019. Studies regarding the diagnostic performance of68Ga-PSMA PET/MRI in detecting primary PCa and biochemical recurrence (BCR) after definitive treatment were included. The quality of each study was assessed using the Quality Assessment of Diagnostic Accuracy Studies-2 (QUADAS-2) tool. The pooled sensitivity and specificity of PET/MRI in identifying primary PCa and the pooled detection rate of PET/MRI for BCR were calculated using a random-effects model. RESULTS: A total of 13 studies with 707 patients were included in the analysis, and the pooled sensitivity and specificity of PET/MRI in detecting primary PCa were 0.83 (95 % CI, 0.73-0.90) and 0.81 (95 % CI, 0.61-0.93), respectively. In the pooled analysis of BCR, the pooled detection rate was 76 % (95 % CI, 72 %-79 %). For four levels of PSA (0-0.2 ng/mL, 0.2-1 ng/mL, 1-2 ng/mL and more than 2 ng/mL), the pooled detection rates were 38 %, 67 %, 74 %, and 95 %, respectively. There was no distinct publication bias, but there was significant study heterogeneity. CONCLUSIONS: 68Ga-PSMA PET/MRI is likely an effective imaging method in the diagnosis of primary PCa. In addition, the diagnostic accuracy of 68Ga-PSMA PET/MRI in patients with BCR was also high, positively correlating with PSA levels.