Spheroidal cancer cell cultures have been used to enrich cancer stem cells (CSC), which are thought to contribute to important clinical features of tumors. This study aimed to map the regulatory networks driven by circular RNAs (circRNAs) in CSC-enriched colorectal cancer (CRC) spheroid cells. The spheroid cells established from two CRC cell lines acquired stemness properties in pluripotency gene expression and multi-lineage differentiation capacity. Genome-wide sequencing identified 1503 and 636 circRNAs specific to the CRC parental and spheroid cells, respectively. In the CRC spheroids, algorithmic analyses unveiled a core network of mRNAs involved in modulating stemness-associated signaling pathways, driven by a circRNA–microRNA (miRNA)–mRNA axis. The two major circRNAs, hsa_circ_0066631 and hsa_circ_0082096, in this network were significantly up-regulated in expression levels in the spheroid cells. The two circRNAs were predicted to target and were experimentally shown to down-regulate miR-140-3p, miR-224, miR-382, miR-548c-3p and miR-579, confirming circRNA sponging of the targeted miRNAs. Furthermore, the affected miRNAs were demonstrated to inhibit degradation of six mRNA targets, viz. ACVR1C/ALK7, FZD3, IL6ST/GP130, SKIL/SNON, SMAD2 and WNT5, in the CRC spheroid cells. These mRNAs encode proteins that are reported to variously regulate the GP130/Stat, Activin/Nodal, TGF-β/SMAD or Wnt/β-catenin signaling pathways in controlling various aspects of CSC stemness. Using the CRC spheroid cell model, the novel circRNA–miRNA–mRNA axis mapped in this work forms the foundation for the elucidation of the molecular mechanisms of the complex cellular and biochemical processes that determine CSC stemness properties of cancer cells, and possibly for designing therapeutic strategies for CRC treatment by targeting CSC.

Vaccination is a key public health strategy that is known to be effective in mitigating the risk of infection and severe disease. However, in the context of the COVID-19 pandemic, the percentage (<50%) of Malaysians who have received a booster for the COVID-19 vaccine has remained stagnant over a year. This study aimed to determine the prevalence of and the factors associated with hesitancy toward the second dose of booster for the COVID-19 vaccine. A web-based cross-sectional study was conducted from August to November 2022. The Oxford Vaccine Hesitancy Scale was used to assess the hesitancy toward the second dose of booster for the COVID-19 vaccine. Simple and multiple factors logistic regressions were used to determine the predictors of hesitancy. A p-value less than 0.05 was considered to be statistically significant. Data from 798 respondents were included in the analysis. The prevalence of hesitancy toward the second booster of the COVID-19 vaccine was 26.7%. The predictors of second-booster hesitancy were older age (AOR = 1.040, 95 CI = 1.022, 1.058), having received the third dose (first booster) because of instruction by the government (AOR = 2.125, 95% CI = 1.380, 3.274), concern about serious long term side effects of the vaccine (AOR = 4.010, 95% CI = 2.218, 7.250), and opinions of close friends and immediate family members that the booster is harmful (AOR = 2.201, 95% CI = 1.280, 3.785). Conversely, factors that appear to reduce vaccine booster hesitancy were acceptance of the third dose due to the high number of cases and the increasing rate of infection (AOR = 0.548, 95% CI = 0.317, 0.947), the belief that the vaccine will decrease the risk of getting the infection (AOR = 0.491, 95% CI = 0.277, 0.870), and opinions of close friends and immediate family members that the booster is helpful (AOR = 0.479, 95% CI = 0.273, 0.840). In conclusion, more than one-fifth of Malaysians were hesitant to take the second booster of the COVID-19 vaccine. This suggests that appropriate steps that increase vaccine acceptance, taking into consideration the findings of the present study, are needed to address this issue and to foster more positive attitudes toward vaccination. The survey was available in three main languages but limited to people with internet access; hence, it would likely be biased toward younger adults and social media users and exclude those with limited or no internet access, in particular older people. Therefore, the results are not representative of the Malaysian population at large and caution should be exercised when interpreting the findings.

Breast cancer, a highly heterogeneous malignancy and a leading cause of cancer-related mortality among women worldwide, is profoundly shaped in its initiation, progression, and therapeutic response by the tumor immune microenvironment (TIME). This review consolidates recent advances in deciphering the cellular, molecular, and metabolic complexity of breast cancer TIME and highlights mechanisms of immune suppression that impede durable treatment efficacy. We critically appraise current and emerging immunotherapeutic approaches, with a focus on strategies that aim to transform immunologically “cold” tumors into “hot,” immune-responsive phenotypes. Novel directions, including metabolic modulation, stromal reprogramming, and precision combination therapies, are discussed in the context of overcoming primary and acquired resistance. We highlight emerging biomarker strategies that integrate spatial transcriptomics to map immune exclusion zones, along with liquid biopsy monitoring of exosomal PD-L1 and circulating tumor DNA, to enable real-time adaptation of combination immunotherapy regimens.

The elderly are considered a high-risk group for severe outcomes and death from COVID-19 infection. Given the emergence of new COVID variants and the immunity provided by vaccines waning over time, booster doses of the vaccine have been advocated for those at risk to stay protected. This study aimed to determine the factors associated with hesitancy toward the second booster of the COVID-19 vaccine among the elderly residing in residential care homes. A cross-sectional study was conducted in 24 residential care homes in the Klang Valley using a face-to-face interview questionnaire. The study population included individuals aged 60 and above who had been fully vaccinated against COVID-19 up to the first booster dose. Second-booster hesitancy was assessed using the Oxford Vaccine Hesitancy Scale with seven items, the aggregate score of which ranges from seven to thirty-five; the higher the score, the greater the level of hesitancy. Multivariate linear regression was employed to determine factors associated with second-booster hesitancy, and a p-value < 0.05 was considered statistically significant. Data from 401 elderly individuals were included for analysis. The mean score of the Oxford Vaccine Hesitancy Scale was 21.6 ± 7.2. Predictors of second booster hesitancy were identified. Age, Indian ethnicity, being a recipient of the Sinovac vaccine as the first COVID-19 booster, experiencing the death of close friends or immediate family members following COVID-19 vaccination, and negative messages (indicating that taking a booster dose is harmful) from caregivers, friends, or family members were found to be associated with an increased second-booster-hesitancy score. Conversely, positive messages (indicating that taking a booster is helpful) from the government and caregivers, friends, or family members were identified as predictors associated with a reduction in the second-booster-hesitancy score. While vaccines effectively combat severe COVID-19, the majority of the elderly hesitate before taking the second booster. Their hesitancy, rooted in the perception of a low self risk and reliance on protection from the initial doses, emphasizes the need for intervention by relevant bodies. Taking into consideration the risk, albeit relatively low, of potentially serious side effects following COVID-19 vaccinations, it is imperative that transparent, appropriate, and positive messaging regarding booster vaccines, particularly in the context of the elderly from residential care homes, be available. Encouraging this high-risk group to embrace the second booster aligns with the goal of maximizing protection within the vulnerable elderly population.

Background: Neoadjuvant immune checkpoint inhibitors combined with chemotherapy improve pathological complete response in early-stage triple-negative breast cancer, but the consistency of benefit, role of PD-L1 expression, and immune-related toxicity remain clinically relevant. Methods: PubMed, Embase, and the Cochrane Central Register of Controlled Trials were searched from inception to June 30, 2026, for randomized trials comparing neoadjuvant immune checkpoint inhibitors plus chemotherapy with chemotherapy alone or placebo plus chemotherapy in early-stage triple-negative breast cancer. The primary efficacy analysis was restricted to phase III trials. Risk ratios and 95% confidence intervals were pooled using a random-effects model. Risk of bias and certainty of evidence were assessed using RoB 2 and GRADE. Results: Four phase III trials including 3,377 patients were included in the primary efficacy synthesis. Neoadjuvant immune checkpoint inhibitors plus chemotherapy significantly increased pathological complete response compared with control treatment (risk ratio, 1.27; 95% confidence interval, 1.19–1.36; P < 0.001), with no statistical heterogeneity. Benefit was observed in both PD-L1-positive tumors (risk ratio, 1.32; 95% confidence interval, 1.18–1.48) and PD-L1-negative tumors (risk ratio, 1.45; 95% confidence interval, 1.22–1.72). Grade ≥ 3 treatment-related adverse events were not significantly increased (risk ratio, 1.06; 95% confidence interval, 0.98–1.13), whereas immune-related adverse events were more frequent with immune checkpoint inhibitors (risk ratio, 3.52; 95% confidence interval, 1.88–6.57). Conclusions: Neoadjuvant immune checkpoint inhibitors plus chemotherapy improve pathological complete response in early-stage triple-negative breast cancer, including across PD-L1-defined subgroups, but increase immune-related toxicity. PD-L1 expression alone appears insufficient for treatment selection. Systematic review registration: https://www.crd.york.ac.uk/PROSPERO/recorddashboard# , identifier CRD420261412366.