B Cells and IL-21-Producing Follicular Helper T Cells Cooperate to Determine the Dynamic Alterations of Premetastatic Tumor Draining Lymph Nodes of Breast Cancer.

Metastasis is the major cause of cancer-related death, and lymph node is the most common site of metastasis in breast cancer. However, the alterations that happen in tumor-draining lymph nodes (TDLNs) to form a premetastatic microenvironment are largely unknown. Here, we first report the dynamic changes in size and immune status of TDLNs before metastasis in breast cancer. With the progression of tumor, the TDLN is first enlarged and immune-activated at early stage that contains specific antitumor immunity against metastasis. The TDLN is then contracted and immunosuppressed at late stage before finally getting metastasized. Mechanistically, B and follicular helper T (Tfh) cells parallelly expand and contract to determine the size of TDLN. The activation status and specific antitumor immunity of CD8+ T cells in the TDLN are determined by interleukin-21 (IL-21) produced by Tfh cells, thus showing parallel changes. The turn from activated enlargement to suppressed contraction is due to the spontaneous contraction of germinal centers mediated by follicular regulatory T cells. On the basis of the B-Tfh-IL-21-CD8+ T cell axis, we prove that targeting the axis could activate TDLNs to resist metastasis. Together, our findings identify the dynamic alterations and regulatory mechanisms of premetastatic TDLNs of breast cancer and provide new strategies to inhibit lymph node metastasis.

Long non-coding RNAFOXD1-AS1 modulated CTCs epithelial-mesenchymal transition and immune escape in hepatocellular carcinoma in vitro by sponging miR-615-3p.

BACKGROUND: Hepatocellular carcinoma (HCC) is widely recognized as a globally prevalent malignancy. Immunotherapy is a promising therapy for HCC patients. Increasing evidence suggests that lncRNAs are involved in HCC progression and immunotherapy. AIM: The study reveals the mechanistic role of long non-coding RNA (lncRNA) FOXD1-AS1 in regulating migration, invasion, circulating tumor cells (CTCs), epithelial-mesenchymal transition (EMT), and immune escape in HCC in vitro. METHODS: This study employed real-time PCR (RT-qPCR) to measure FOXD1-AS1, miR-615-3p, and programmed death-ligand 1 (PD-L1). The interactions of FOXD1-AS1, miR-615-3p, and PD-L1 were validated via dual-luciferase reporter gene and ribonucleoprotein immunoprecipitation (RIP) assay. In vivo experimentation involves BALB/c mice and BALB/c nude mice to investigate the impact of HCC metastasis. RESULTS: The upregulation of lncRNA FOXD1-AS1 in malignant tissues significantly correlates with poor prognosis. The investigation was implemented on the impact of lncRNA FOXD1-AS1 on the migratory, invasive, and EMT of HCC cells. It has been observed that the lncRNA FOXD1-AS1 significantly influences the generation and metastasis of MCTC in vivo analysis. In mechanistic analysis, lncRNA FOXD1-AS1 enhanced immune escape in HCC via upregulation of PD-L1, which acted as a ceRNA by sequestering miR-615-3p. Additionally, lncRNA FOXD1-AS1 was found to modulate the EMT of CTCs through the activation of the PI3K/AKT pathway. CONCLUSION: This study presents compelling evidence supporting the role of lncRNA FOXD1-AS1 as a miRNA sponge that sequesters miR-655-3p and protects PD-L1 from suppression.

Axilla View of Mammography in Preoperative Axillary Lymph Node Evaluation of Breast Cancer Patients: A Pilot Study.

PURPOSE: This study aimed to explore a novel position of mammography named axilla view in axillary lymph node (ALN) evaluation in breast cancer. PATIENTS AND METHODS: Patients were prospectively enrolled and scheduled for mammography before surgery. Investigated imaging patterns included mediolateral oblique (2D-MLO) and axilla view (2D-axilla) of mammography, and axilla view of digital breast tomosynthesis (3D-axilla). The correlation of ALN numbers between imaging and pathology was analyzed. Diagnostic performance was analyzed via AUC. RESULTS: 75 patients were included. A larger and clearer axillary region was displayed in axilla view. The total number of ALNs detected under 2D/3D-axilla view was significantly higher than that under 2D-MLO view (4.6 vs. 2.5, P < .001; 5.6 vs. 4.6, P = .034). Correlations between number of positive ALNs detected under 2D/3D-axilla view and pathologically confirmed metastatic ALNs were stronger than 2D-MLO view (Pearson correlation coefficients: 0.7084,0.7044 and 0.4744). The proportion of cases with ≥5 positive ALNs detected under 3D-axilla view was significantly higher than that under 2D-MLO (38.2% vs. 14.7%, P = .028). The overweight and obese group showed a higher AUC value than the underweight and lean group in ALN evaluation, although not significantly (2D-MLO: 0.7643 vs. 0.6458, P = .2656; 2D-axilla: 0.8083 vs. 0.6586, P = .1522; 3D-axilla: 0.8045 vs. 0.6615, P = .1874). This difference was more pronounced in axilla view. CONCLUSION: Axilla view exhibited advantages over conventional MLO view in the extent of axilla displayed by mammography in breast cancer. Further studies with larger sample sizes are needed.

Development of a combined model incorporating clinical characteristics and magnetic resonance imaging features to enhance the predictive value of a prognostic model for locally advanced cervical cancer.

Objective: This study aimed to develop non-invasive predictive tools based on clinical characteristics and magnetic resonance imaging (MRI) features to predict survival in patients with locally advanced cervical cancer (LACC), thereby facilitating clinical decision-making. Methods: We conducted a retrospective analysis of clinical and MRI data from LACC patients who underwent radical radiotherapy at our center between September 2012 and May 2020. Prognostic predictors were identified using single-factor and multifactor Cox analyses. Clinical and MRI models were established based on relevant features, and combined models were created by incorporating MRI factors into the clinical model. The predictive performance of the models was evaluated using the area under the curve (AUC), consistency index (C-index), and decision curve analysis (DCA). Results: The study included 175 LACC patients. Multivariate Cox analysis revealed that patients with FIGO IIA-IIB stage, ECOG score 0-1, CYFRA 21-1<7.7 ng/ml, ADC ≥ 0.79 mm^2/s, and Kep ≥ 4.23 minutes had a more favorable survival prognosis. The clinical models, incorporating ECOG, FIGO staging, and CYFRA21-1, outperformed individual prognostic factors in predicting 5-year overall survival (AUC: 0.803) and 5-year progression-free survival (AUC: 0.807). The addition of MRI factors to the clinical model (AUC: 0.803 for 5-year overall survival) increased the AUC of the combined model to 0.858 (P=0.011). Similarly, the combined model demonstrated a superior predictive ability for 5-year progression-free survival, with an AUC of 0.849, compared to the clinical model (AUC: 0.807) and the MRI model (AUC: 0.673). Furthermore, the C-index of the clinical models for overall survival and progression-free survival were 0.763 and 0.800, respectively. Upon incorporating MRI factors, the C-index of the combined model increased to 0.826 for overall survival and 0.843 for progression-free survival. The DCA further supported the superior prognostic performance of the combined model. Conclusion: Our findings indicate that ECOG, FIGO staging, and CYFRA21-1 in clinical characteristics, as well as ADC and Kep values in MRI features, are independent prognostic factors for LACC patients undergoing radical radiotherapy. The combined models provide enhanced predictive ability in assessing the risk of patient mortality and disease progression.

Exosome-derived ANXA9 functions as an oncogene in breast cancer.

Breast cancer (BCA) is one of the most prevalent cancers among women. Emerging evidence has revealed that Annexin A-9 (ANXA9) plays a crucial function in the development of some cancers. Notably, ANXA9 has been reported to be a new prognostic biomarker for gastric and colorectal cancers. However, its expression and biological function in BCA have not yet been investigated. Using online bioinformatics tools such as TIMER, GEPIA, HPA, and UALCAN, we predicted ANXA9 expression and its correlation with the clinicopathological characteristics of BCA patients. RT-qPCR and western blot were utilized to measure ANXA9 mRNA and ANXA9 protein expression in BCA patient tissues and cells. BCA-derived exosomes were identified by transmission electron microscopy. Functional assays were employed to evaluate the biological role of ANXA9 in BCA cell proliferation, migration, invasion, and apoptosis. A tumor xenograft in vivo model was utilized to assess the role of ANXA9 in tumor growth in mice. Bioinformatics and functional screening analysis revealed that ANXA9 was highly expressed in BCA patient tissues, with median ANXA9 expression 1.5- to 2-fold higher than in normal tissues (p < 0.05). RT-qPCR confirmed that ANXA9 expression in BCA tissues was around 1.5-fold higher than the adjacent normal tissues (p < 0.001). ANXA9 expression in different subtypes of BCA also showed a difference, and ANXA9 was found to be mostly significantly upregulated in luminal BCA relative to normal tissues or other histological subtypes (p < 0.001). Moreover, ANXA9 expression was elevated in different races, ages, clinical stages, node metastasis status, and menopause status groups relative to the normal group (p < 0.001). Furthermore, ANXA9 was found to be secreted by BCA tissue-derived exosomes and its expression was upregulated 1- to 7-fold in BCA cells treated with exosomes (p < 0.001), while its expression in MCF10A cells was not significantly altered by treatment with exosomes (p > 0.05). ANXA9 silencing induced a significant decrease of around 30% in the colony number of BCA cells (p < 0.01). The number of migrated and invaded BCA cells also decreased by around 65 and 68%, respectively, after silencing ANXA9 (p < 0.01). Tumor size was significantly reduced (nearly half) in the LV-sh-ANXA9 group relative to the LV-NC group in the xenograft model (p < 0.01), suggesting that ANXA9 silencing repressed tumor progression in BCA progression in vitro and in vivo. In conclusion, exosome-derived ANXA9 functions as an oncogene that facilitates the proliferation, migration, and invasiveness of BCA cells and enhances tumor growth in BCA development, which may provide a new prognostic and therapeutic biomarker for BCA patients.

HER2-low status may predict poor neoadjuvant chemotherapy response in HR-negative breast cancer: a real-world multicenter study.

OBJECTIVE: We aimed to investigate the impact of human epidermal growth factor receptor 2 status (human epidermal growth factor receptor 2-low versus human epidermal growth factor receptor 2-zero) on pathological response to neoadjuvant chemotherapy and survival outcomes in early-stage breast cancer. METHODS: Patients with primary invasive breast cancer received neoadjuvant chemotherapy between July 2018 and July 2021 were identified from six hospitals. The primary efficacy end-point was total pathological complete response. The second short-term efficacy end-points include breast pathological complete response, axillary lymph nodes pathological complete response and the score of Miller-Payne grade. Long-term efficacy end-point was disease-free survival. RESULTS: 429 patients with human epidermal growth factor receptor 2 negative invasive tumors were included, 267 (62.24%) had human epidermal growth factor receptor 2-low tumors. Hormone receptor-positive patients had a higher percentage of human epidermal growth factor receptor 2-low tumors compared to hormone receptor-negative patients (71.97% versus 42.14%). The pathological response rate was significantly lower in human epidermal growth factor receptor 2-low tumors than in human epidermal growth factor receptor 2-zero tumors for total patients in univariate analysis, including the rates of total pathological complete response (5.2% versus 14.2%), breast pathological complete response (6.4% versus 17.3%), nodes pathological complete response (26.3% versus 37.7%) and MP4-5 (21.2% versus 33.8%). Subgroup analysis showed that the rates of total pathological complete response, breast pathological complete response and MP4-5 were also significantly lower in human epidermal growth factor receptor 2-low tumors versus human epidermal growth factor receptor 2-zero tumors in both univariate and multivariate analysis in hormone receptor-negative subgroup. With the median follow-up of 24 months, disease-free survival was comparable between these two subgroups (P = 0.816). CONCLUSIONS: Our results demonstrate that human epidermal growth factor receptor 2-low tumors achieved a significantly lower pathological complete response rate with conventional chemotherapy than those with human epidermal growth factor receptor 2-zero tumors, especially for hormone receptor-negative group. Large, randomized, prospective studies are needed to confirm our data and further evaluate the prognostic value of human epidermal growth factor receptor 2-low expression.

The emerging role of adopting protamine for reducing the risk of bleeding complications during the percutaneous coronary intervention: A meta-analysis.

BACKGROUND: The safety and the benefits of reducing the risk of bleeding complications via protamine administration during the percutaneous coronary intervention (PCI) remains unclear. This study aimed to systematically assessed the efficacy and safety of using protamine in PCI. METHOD: Potential academic studies were identified from PubMed, Cochrane Library, EMBASE, and Web of Science. The time range we retrieved from was that from the inception of electronic databases to March 31, 2022. Gray studies were identified from the references of included literature reports. Stata version 12.0 statistical software (StataCorp LP) was used to analyze the pooled data. RESULTS: A total of seven studies were involved in our study. The overall participants of the protamine group were 4983, whereas it was 1953 in the nonprotamine group. This meta-analysis indicated that protamine was preferable for PCI as its lower value of major bleeding (odds ratio [OR] = 0.489, 95% confidence interval [CI]: 0.362-0.661, p < .001) and minor bleeding (OR = 0.281, 95% CI: 0.123-0.643, p = .003). Additionally, the protamine did not tend to be related a higher incidence of mortality (p = .143), myocardial infarction (p = .990), and stent thrombosis (p = .698). CONCLUSIONS: Based on available evidence, use of protamine may reduce the risk of bleeding complications without increasing the risk of mortality, myocardial infarction, and stent thrombosis. Given the relevant possible biases in our study, adequately powered and better-designed studies with long-term follow-up are required to reach a firmer conclusion.

Ultrasound-Guided Percutaneous Transhepatic Gallbladder Drainage Improves the Prognosis of Patients with Severe Acute Cholecystitis.

The aim of this study was to investigate the clinical efficacy of ultrasound-guided percutaneous transhepatic gallbladder drainage (PTGD) for the treatment of severe acute cholecystitis (AC). The data of 40 patients diagnosed with severe AC at our hospital between August 2020 and June 2021 were retrieved and classified into a PTGD group, open cholecystostomy (OC) group, laparoscopic cholecystectomy (LC) group, and conventional conservative treatment (CT) group. Before treatment and on days 1, 3, 5, and 7 after treatment, their serum levels of alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), total bilirubin (TBIL), triglyceride (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), white blood cell count (WBC), IL-2, IL-4, IL-6, IL-8, and cancer antigen 19-9 (CA19-9) were measured. Additionally, clinical manifestations such as body temperature and pain score were monitored before treatment and at 24, 48, and 72 hours after treatment. The recovery time and complications/adverse reactions were statistically analyzed, and the Kaplan-Meier survival curve was plotted. After treatment, compared with the other three groups, the PTGD group had a significant reduction in serum indicators, including WBC and inflammatory factors, recovery time, pain score, and complications, and benefitted from better treatment efficacy and higher survival rate. Thus, ultrasound-guided PTGD was found to be more effective in treating severe AC patients and was associated with improved patient prognoses.

Local and systemic therapy may be safely de-escalated in elderly breast cancer patients in China: A retrospective cohort study.

Background: For elderly patients with breast cancer, the treatment strategy is still controversial. In China, preoperative axillary lymph node needle biopsy is not widely used, resulting in many patients receiving axillary lymph node dissection (ALND) directly. Our study aims to determine whether local and systemic therapy can be safely de-escalated in elderly breast cancer. Methods: Patients aged ≥70 years were retrospectively enrolled from our institution's medical records between May 2013 and July 2021. Groups were assigned according to local and systemic treatment regimens, and stratified analysis was performed by molecular subtypes. Univariate and multivariate survival analyses were used to compare the effects of different regimens on relapse-free survival (RFS). Results: A total of 653 patients were enrolled for preliminary data analysis, and 563 patients were screened for survival analysis. The mean follow-up was 19 months (range, 1-82 months). Axillary lymph node metastases were pathologically confirmed in only 2.1% of cN0 cases and up to 97.1% of cN+ cases. In the aspect of breast surgery, RFS showed no significant difference between mastectomy and BCS group (p = 0.3078). As for axillary surgery, patients in the ALND group showed significantly better RFS than those in the sentinel lymph node biopsy (SLNB) group among pN0 patients (p = 0.0128). Among these cases, the proportion of cN+ in ALND was significantly higher than that in SLNB (6.4% vs. 0.4%, p = 0.002), which meant axillary lymph nodes (ALNs) of ALND patients were larger in imaging and more likely to be misdiagnosed as metastatic. With regard to adjuvant therapy, univariate and multivariate analyses showed that RFS in different comprehensive adjuvant regimens were similar especially among hormone receptor (HR)+/human epidermal growth factor receptor 2 (HER2)- subgroup where patients who did not receive any adjuvant therapy accounted for 15.7% (p > 0.05). Conclusions: It is feasible to reduce some unnecessary local or systemic treatments for elderly breast cancer patients, especially in HR+/HER2- subtype. Multiple patient-related factors should be considered when making treatment plans.

Evaluation of short- and medium-term efficacy and complications of ultrasound-guided ablation for small liver cancer.

BACKGROUND: To ensure clinical efficacy and prolong patient survival, treatments such as surgery and microwave ablation (MWA) are used for early liver cancer. MWA is preferred because it effectively preserves the normal liver tissue and causes transient coagulation necrosis of local liver tumor cells. However, due to technical limitations, the cancerous liver tissue cannot be completely ablated; therefore, the probability of local tumor recurrence is high. AIM: To investigate the clinical efficacy and safety of ultrasound-guided percutaneous MWA in the treatment of small liver cancer. METHODS: A total of 118 patients treated for small liver cancer in The Central Hospital of Yongzhou from January 2018 to April 2019 were selected. Sixty-six patients received ultrasound-guided percutaneous MWA (MWA group) and 52 received laparoscopic surgery (laparoscope group). The operation time, blood loss, hospital stay, and medical expenses of both groups were statistically analyzed. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), albumin (ALB), alpha fetal protein (AFP), carcinoembryonic antigen (CEA), and peripheral blood regulatory T lymphocytes (Treg) levels were evaluated pre- and post-operatively. The cross-sectional area of tumors measured before and after ablation was analyzed statistically; the therapeutic effect was compared between both groups in terms of surgical complications, 2-year progression-free survival rate, and overall survival rate. RESULTS: The operation time, blood loss, hospital stay, and medical expenses in the MWA group were lower than those of the laparoscope group, and the differences were significant (P < 0.05); these parameters, and ALT, AST, TBIL, and ALB levels were compared preoperatively between both groups, and there was no significance (P > 0.05). The operation time, blood loss, hospital stay, and medical expenses for 2 d and 1 wk after surgery, the ALT and AST of the MWA group were lower than those of the laparoscope group, and the difference was significant (P < 0.05). The operation time, blood loss, hospital stay, and medical expenses, and serum AFP, CEA, and Treg levels were measured preoperatively and 4 and 8 wk postoperatively, and there were no significant differences between the two groups (P > 0.05). Compared with preoperative levels, serum AFP, CEA, and Treg levels in both groups were decreased (P < 0.05). The lesion in the MWA group had a maximum area of 4.86 ± 0.90 cm2, 1.24 ± 0.57 cm2, and 0.31 ± 0.11 cm2 preoperatively, 1 and 3 mo postoperatively, respectively. Fifty-eight of them achieved complete response and eight achieved a partial response. After 2 years of follow-up, the progression-free and overall survival rates in the MWA group were 37.88% and 66.67%, respectively, compared with 44.23% and 76.92% in the laparoscope group, with no significant difference (P > 0.05). CONCLUSION: The effects of ultrasound-guided percutaneous MWA in the treatment of small liver cancer are similar to those of laparoscopic surgery. However, ablation causes less trauma and liver dysfunction.

Long non-coding RNA CCL2 promoted gastric cancer function via miR-128/ PARP2 signal pathway.

Amounts of studies have revealed long non-coding RNA (lncRNA) was related to the development of gastric cancer. Here, our results suggested the function and regulatory mechanism of CCL2 in gastric cancer. Quantitative polymerase-chain reaction (qPCR) was employed to inspect lncRNA CCL2 and miR-128 expression in normal gastric cell line (GES-1) and tumor cell lines (HGC-27 and MKN-45). The effects of CCL2 and miR-128 were measured via Luciferase reporter test. Western blot was used to check PARP2 protein expression. CCL2 expression and PARP2 protein levels were up-regulated, while miR-128 expression was obviously lower. Meanwhile, CCL2 down-regulating significantly repressed the proliferation, migration, and invasion by regulating miR-128. In addition, we proved miR-128 was a direct target of CCL2 through double luciferase assay and bioinformatics analysis. Moreover, miR-128 markedly inhibited the proliferation, migration, and invasion in gastric cancer. More importantly, miR-128 could reverse the effects of lncRNA CCL2 knocked down. PARP2-si obviously suppressed in gastric cancer proliferation, migration, and invasion. Meanwhile, miR-128 mimic and the knockout of CCL2 distinctly decreased PARP2 protein level. Additionally, luciferase report experiments certificated that PARP2 targeted miR-128, implying PARP2 directly interacted with miR-128 in gastric cancer. More interestingly, the downregulation of PARP could reverse the trend triggered by miR-128 inhibitor in gastric tumor. All over these results showed lncRNA CCL2 played importance of role in gastric tumor via miR-128/PARP2 axis signal pathway. LncRNA CCL2 accelerated gastric cancer progression by regulating miR-128/PARP2 signaling pathway, providing a novel possible strategy for the treatment of gastric cancer.

[Molecular genetic study of a family featuring cardiac conduction block].

OBJECTIVE To explore the genetic mechanism for a family affected with cardiac conduction block. METHODS Affected family members were screened for potential mutations of known candidate genes. As no pathogenic mutation was found, two patients and one healthy member from the family were further analyzed by exomic sequencing followed by Sanger sequencing. The pathogenicity of suspected mutation was analyzed using bioinformatics software. RESULTS Sequencing of the full exome has identified a c.G1725T mutation in the CLCA2 gene. Sanger sequencing has detected the same mutation in all five patients, but not in the normal member from the family. Bioinformatics analysis indicated that the mutation has resulted in substitution of the 575th amino acid cysteine (C) by tryptophan (W). The site is highly conserved and becomes pathogenic with the mutation. CONCLUSION The heterozygous c.G1725T mutation in exon 11 of the CLCA2 gene probably underlies the disease and fit the autosomal dominant pattern of inheritance.