Establishment of a cholangiocarcinoma risk evaluation model based on mucin expression levels.

BACKGROUND: Cholangiocarcinoma (CCA) is a highly malignant cancer, characterized by frequent mucin overexpression. MUC1 has been identified as a critical oncogene in the progression of CCA. However, the comprehensive understanding of how the mucin family influences CCA progression and prognosis is still incomplete. AIM: To investigate the functions of mucins on the progression of CCA and to establish a risk evaluation formula for stratifying CCA patients. METHODS: Single-cell RNA sequencing data from 14 CCA samples were employed for elucidating the roles of mucins, complemented by bioinformatic analyses. Subsequent validations were conducted through spatial transcriptomics and immunohistochemistry. The construction of a risk evaluation model utilized the least absolute shrinkage and selection operator regression algorithm, which was further confirmed by independent cohorts and diverse data types. RESULTS: CCA tumor cells with elevated levels of MUC1 and MUC4 showed activated nucleotide metabolic pathways and increased invasiveness. MUC5AC-high cells were found to promote CCA progression through WNT signaling. MUC5B-high cells exhibited robust cellular oxidation activities, leading to resistance against antitumoral treatments. MUC13-high cells were observed to secret chemokines, recruiting and transforming macrophages into the M2-polarized state, thereby suppressing antitumor immunity. MUC16-high cells were found to promote tumor progression through interleukin-1/nuclear factor kappa-light-chain-enhancer of activated B cells signaling upon interaction with neutrophils. Utilizing the expression levels of these mucins, a risk factor evaluation formula for CCA was developed and validated across multiple cohorts. CCA samples with higher risk factors exhibited stronger metastatic potential, chemotherapy resistance, and poorer prognosis. CONCLUSION: Our study elucidates the functional mechanisms through which mucins contribute to CCA development, and provides tools for risk stratification in CCA.

SPINK1 Overexpression Correlates with Hepatocellular Carcinoma Treatment Resistance Revealed by Single Cell RNA-Sequencing and Spatial Transcriptomics.

Low efficacy of treatments and chemoresistance are challenges in addressing refractory hepatocellular carcinoma (HCC). SPINK1, an oncogenic protein, is frequently overexpressed in many HCC cases. However, the impact of SPINK1 on HCC treatment resistance remains poorly understood. Here, we elucidate the functions of SPINK1 on HCC therapy resistance. Analysis of SPINK1 protein level reveals a correlation between elevated SPINK1 expression and unfavorable prognosis. Furthermore, intercellular variations in SPINK1 expression levels are observed. Subsequent examination of single cell RNA-sequencing data from two HCC cohorts further suggest that SPINK1-high cells exhibit heightened activity in drug metabolic pathways compared to SPINK1-low HCC cells. High SPINK1 expression is associated with reduced sensitivities to both chemotherapy drugs and targeted therapies. Moreover, spatial transcriptomics data indicate that elevated SPINK1 expression correlates with non-responsive phenotype during treatment with targeted therapy and immune checkpoint inhibitors. This is attributed to increased levels of drug metabolic regulators, especially CES2 and CYP3A5, in SPINK1-high cells. Experimental evidence further demonstrates that SPINK1 overexpression induces the expression of CES2 and CYP3A5, consequently promoting chemoresistance to sorafenib and oxaliplatin. In summary, our study unveils the predictive role of SPINK1 on HCC treatment resistance, identifying it as a potential therapeutic target for refractory HCC.

Molecular pathology assists the diagnosis of lymphoepithelial sialadenitis, Sjögren's syndrome and extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue.

Background/purpose: Lymphoepithelial sialadenitis (LESA), Sjögren's syndrome (SS), and salivary MALT lymphoma are diseases characterized by lymphoepithelial lesions, and the differential diagnosis between them in the salivary glands is challenging. This study aimed to explore clinicopathological and genetic characteristics of the three diseases. Materials and methods: We retrospectively analyzed the clinical features, the histomorphology, immunohistochemistry, and genetic profiling by polymerase chain reaction (PCR) and next-generation sequencing (NGS). Results: There included 68 LESAs, 25 SSs, and 62 MALT lymphomas. Ten cases relapsed in total, and 3 of MALT lymphomas died due to high-level transformation. Immunohistochemical double staining showed FCRL4 cells co-expressed Pax-5 and Ki-67, suggesting FCRL4 cells were proliferative B-cells. The expression level of the FCRL4 was significantly higher in MALT lymphoma than LESA and SS. The detection rates of clonal IGH, IGK, and IGL gene rearrangements in MALT lymphoma with a sensitivity of 83.33%. Monoclonal immunoglobulin gene rearrangements were confirmed in five suspected patients by NGS (100%). Conclusion: FCRL4 B cells might play an important role in the formation of lymphoepithelial lesions and might be as a diagnostic positive marker of salivary MALT lymphoma. The application of multiple detection methods could significantly improve the diagnostic accuracy for MALT lymphomas from LESA and SS.

The impact of neoadjuvant chemotherapy on low anterior resection syndrome after rectal cancer resection: A 6 Months longitudinal follow-up.

BACKGROUND/OBJECTIVE: Neoadjuvant radiotherapy plays a vital role in rectal cancer treatment, but impairs postoperative bowel function, leading to low anterior resection syndrome (LARS). Neoadjuvant chemotherapy alone might avoid the negative effect of radiotherapy on bowel function. This study aims to assess the impact of neoadjuvant chemotherapy on LARS and the development of LARS over the first 6 months after surgery. METHODS: Rectal cancer patients were prospectively recruited during June 30, 2018 and December 24, 2019. Bowel function was assessed by the LARS score, which was taken at 1 month, 3 months, and 6 months after surgery via phone call interview. Patients were divided into two groups based on whether they received neoadjuvant chemotherapy (group A) or not (group B). RESULTS: A total of 97 patients were included in the analysis. There was no significant difference between the LARS scores at 1 month, 3 months, and 6 months of both groups. The LARS score at 6 months showed a significant decrease from that of 1 month and 3 months in group B (P < 0.05, P < 0.01) and in all patients (P < 0.05, P = 0.001), and significant difference was found between the LARS scores in group A at the three timepoints (P < 0.05). No significant difference was found between the scores at 1 month and 3 months in both groups and in all patients. CONCLUSION: Neoadjuvant chemotherapy alone did not have a negative impact on LARS. The bowel function after surgery started to show significant improvement at 6 months after surgery.

Pectolinarigenin flavonoid exhibits selective anti-proliferative activity in cisplatin-resistant hepatocellular carcinoma, autophagy activation, inhibiting cell migration and invasion, G2/M phase cell cycle arrest and targeting ERK1/2 MAP kinases.

PURPOSE: The main purpose of the present research article was to investigate the anticancer properties of pectolinarigenin flavonoid in cisplatin-resistant hepatocellular carcinoma cells (SK-HEP-1) and normal liver cells (AML-12), along with examining its effects on autophagy, cell migration and invasion, cell cycle arrest and ERK1/2 MAP signalling pathways. METHODS: Antiproliferative effects in cancer and normal cells were assessed by MTT cell viability assay. Cell autophagy effects were studied by electron microscopy as well as western blot. Effects on cell cycle were evaluated by flow cytometry using Annexin V/propidium iodide (PI) staining. Transwell migration assay and in vitro wound healing assay were performed to study the effects on cell migration and invasion, respectively. RESULTS: The results indicated that pectolinarigenin inhibited significantly the growth of the SK-HEP-1 liver cancer cells and exhibited an IC50 of 10 µM, while against normal cells the cytotoxic effects were much less pronounced. Further, it was observed that the anticancer effects of pectolinarigenin were due to induction of autophagy which was also associated with upregulation of the expression of Beclin-1, LC3-I and LC3-II. Transmission electron microscopy showed the formation of autophagosomes and vesicles. Pectolinarigenin also caused arrest of the SK-HEP-1 cells at the G2/M-phase of the cell cycle. Wound healing and transwell assays showed pectolinarigenin suppressed the migration and invasive potential of the SK-HEP-1 cells. CONCLUSIONS: The present study revealed that pectolinarigenin exhibits antitumor activity in SK-HEP-1 liver cancer cells via multiple mechanisms and may prove promising in the development of systemic therapy for liver cancer.

[The clinical efficacy of 0.1% bromfenac sodium hydrate ophthalmic solution after excimer laser in situ keratomileusis].

OBJECTIVE: To investigate the clinical efficacy and safety of 0.1% bromfenac sodium hydrate ophthalmic solution in myopia and astigmatism eyes after sub-Bowman keratomileusis (SBK) METHODS: A case control study. Number of patients with low to moderate myopia (-6.00 D ≤ spherical equivalent < -2.00 D) in the test and control groups was 17 cases (32 eyes) and 20 cases (40 eyes), respectively. Number of patients with high myopia (-11.00 D ≤ spherical equivalent < -6.00 D) in the test and control groups was 22 cases (42 eyes) and 15 cases (26 eyes) respectively. The first day after SBK, 0.1% bromfenac sodium hydrate eye drops was administrated 2 times/day in patients in the test group and continued for 10 and 14 days in low and high myopia, respectively. In the control group, 0.1% fluorometholone eye drops was used 4 times/day, then reduced gradually and continuously for 16 days. Visual acuity, computer refraction, intraocular pressure (IOP) and corneal topography examination were conducted at different postoperative period. Symptoms and related complications were recorded. Ranked data were statistically analyzed using the Wilcoxon rank sum test and quantitative data were analyzed using independent samples t-test. RESULTS: For low to moderate myopic patients, average postoperative IOP and corneal curvature K2 after one and three months in the test group were (7.84 ± 1.35) and (8.13 ± 1.75) mm Hg(1 mm Hg = 0.133 kPa), and (38.66 ± 1.68) and (38.75 ± 1.45) D, respectively; in the control group, these parameters were (9.37 ± 1.28) and (9.47 ± 1.58) mm Hg and (39.56 ± 1.58), and (39.51 ± 1.50) D, respectively. All of these data in the test group were lower than those in the control group, the differences were statistically significant (t = -2.299, -2.112, P < 0.05). There were no significant differences (P < 0.05) in postoperative uncorrected visual acuity, spherical equivalent, corneal curvature K1, astigmatism CY, IOP (after 6 months) and corneal curvature K2 (after 6 months) between the test and control group. For the high myopic patients, mean postoperative uncorrected visual acuity in the test group and control group after 3 months were 5.14 ± 0.06 and 5.09 ± 0.07, respectively. The difference was statistically significant (t = 2.517, P = 0.015). There were no significant differences in symptoms between these two groups (two-sided test P > 0.1). High IOP and obvious myopia regression were not found in all patients. CONCLUSIONS: Bromfenac sodium hydrate eye drops (0.1%) can achieve the same therapeutic effect as fluorometholone eye drops after SBK and the former is better than the later in postoperative IOP control.