Fecal microbiota transplantation (FMT) has advanced significantly as a therapeutic approach over the past few decades. Preparing fecal suspensions for FMT is one of the key steps. However, there is no unified standard or recognized procedure for preparing fecal suspensions in laboratories. This review evaluated the steps currently employed in laboratories to prepare fecal suspensions for FMT, including sample collection, suspension buffers, homogenization, purification, filtration, centrifugation, cryopreservation, dosage, the operating environment, and the transplantation form. This review focuses on the different operations of each preparation step, aiming to provide a reference for the laboratory preparation of fecal suspensions.

Background: The preparation method of fecal microbiota suspensions is crucial for reliable fecal microbiota transplantation research. However, current protocols vary significantly in processing parameters, potentially compromising the comparability of studies. Systematic comparisons of how different preparation techniques affect the microbial community are still lacking. Methods: Fresh fecal samples from nine adult volunteers were processed via filtration and centrifugation at various speeds. Microbial viability was assessed via live/dead staining and colony forming unit enumeration. 16S rDNA sequencing was performed to analyze bacterial diversity and taxonomic composition. Results: The microbial composition and functional potential did not differ between the filtration alone group and the fresh fecal group, and filtration resulted in the lowest bacterial mortality. Mortality increased with increasing centrifugation speed. Centrifugation selectively affected the abundance of some genera (e.g., Delftia and Acinetobacter ). High-speed supernatants presented markedly lower OD ₆₀₀ values than low-speed supernatants did, and differential centrifugation visibly reduced the amount of debris. Conclusion: Filtration alone best preserved fecal microbial viability, composition and functional potential. Centrifugation enrichment becomes selective at high speeds. Differential centrifugation offered superior impurity removal. The preparation strategy should be tailored to the research or therapeutic goal.

In gastric cancer, gastrokine 1 (GKN1) is a potential theragnostic marker while the related mechanisms remain elusive. Exosomes mediate intercellular communications via transferring various molecules, yet there are limited research studies on the specific cargos of gastric cancer exosomes and the associated mechanisms in this disease. In the present study, AGS and N87‐C cells were transfected with an overexpressed GKN1 plasmid, followed by extraction of exosomes. The study utilized gastric cancer cell lines and a xenograft mouse model to investigate the functional significance of exosomal GKN1. Cell proliferation, metastasis, and apoptosis were assessed through CCK‐8, Transwell, and flow cytometry assays, respectively. The study further explored the mechanism of exosomal GKN1 and its interaction with the PI3K/AKT/mTOR signaling pathways, including immunofluorescence and western blot analyses. Exosomal GKN1 was observed to suppress cell proliferation and invasion while enhancing apoptosis. This effect was attributed to the modulation of key proteins involved in cellular processes, including Ki‐67, MMP‐9, Bcl‐2, Bax, caspase‐3, and caspase‐9, ultimately impacting the PI3K/AKT/mTOR signaling pathway. The findings suggest that exosomal GKN1 exerts inhibitory effects on gastric cancer cell growth and invasion through the regulation of the PI3K/AKT/mTOR signaling cascade, both in experimental cell cultures and animal models.

Smoking is a key driver of esophageal, stomach, liver, pancreatic, and colorectal cancers, and its impact is stronger in adults aged ≥60 years due to cumulative exposure and population aging. A comprehensive analysis of temporal trends and sociodemographic drivers for major digestive cancers in this population is lacking. This study quantified smoking-attributable deaths and disability-adjusted life years (DALYs) from digestive cancers in adults aged ≥60 years globally from 1990 to 2021. It also projected future burdens to 2036 to inform age-focused tobacco control and cancer prevention. This study is a secondary dataset analysis of data from GBD 2021. We analyzed smoking-attributable deaths and DALYs for esophageal, stomach, liver, pancreatic, and colorectal cancers among adults aged ≥60 years across 204 countries and territories (1990–2021). We decomposed temporal changes into contributions from population growth, aging, and changes in age-specific rates. Associations with the sociodemographic index (SDI) were assessed, and future burdens (2022–2036) were projected using a Bayesian age-period-cohort model. From 1990 to 2021, global smoking-attributable deaths increased by 27.5% (398426 to 508147) and DALYs by 14.6% (8.9 to 10.2 million). Decomposition attributed the increase primarily to population growth (41–43%), partially offset by declining age-specific rates (-30% to -35%). Regional disparities were stark: high-SDI regions stabilized burdens through epidemiological improvements, while low-SDI regions saw population-driven escalations. SDI was positively correlated with burden at the country level (ρ=0.20–0.48; p<0.001), peaking at mid-high SDI. Despite falling age-specific rates, the absolute burden of smoking-attributable digestive cancers in older adults is rising in parallel with demographic growth. Projections to 2036 suggest that, if current demographic trends continue, absolute deaths and DALYs are expected to continue increasing as populations age and grow.

Background: Earlier studies have revealed associations between particular ion channel-related genes (ICRGs) and adverse outcomes in gastric cancer (GC). This study aimed to examine the potential relationship between ICRGs and GC. Methods: By integrating differential gene expression analysis of The Cancer Genome Atlas (TCGA) dataset with ICRG sets from GeneCards and performing subsequent regression analysis, an ion channel-related gene risk score (ICRGRS) for GC was identified. We assessed the ability of the ICRGRS to grade the prognosis, gene mutations, pathway enrichment, immunological characteristics, immunotherapy response, and drug sensitivity of GC patients. The relative expressions of hub genes in GC cells were determined using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Cell function assays were used to examine the effect of CNGB3 on the malignant function of GC cells. Results: Eight prognostic ICRGs for GC were selected, leading to the creation of an ICRGRS designed to predict GC outcomes. Datasets from TCGA and Gene Expression Omnibus affirmed the strong forecasting potential of ICRGRS regarding GC survival. Moreover, ICRGRS functioned independently as a determinant of GC prognosis. Utilizing ICRGRS together with clinical variables, a nomogram was formulated to predict survival in GC cases. Evaluations of immune infiltration patterns and drug susceptibility in cohorts stratified by high versus low ICRGRS revealed that elevated ICRGRS instances featured an immunosuppressive tumor microenvironment and displayed superior gains from ZM447439, RO-3306, NU7441, BMS-754807, and JQ1. On the other hand, low-ICRGRS cases showed increased efficacy toward immunotherapies and conventional GC agents like Oxaliplatin and 5-fluorouracil. Through RT-qPCR, heightened CNGB3 transcripts were observed in AGS and HGC27 cells over GES-1. Functional assays indicated that depleting CNGB3 substantially curtailed the proliferation, migration, and invasion of GC cells. Additionally, knockdown of CNGB3 suppressed the intracellular Ca²⁺ concentration and upregulated the expression of major histocompatibility complex class I (MHC-I) in GC cells. Conclusions: Our study established a risk model for predicting the prognosis of GC based on 8 prognostic ICRGs. CNGB3 knockdown significantly suppressed the malignant phenotype of GC cells. Our findings furnish innovative viewpoints on outcome forecasting and therapeutic targets of GC through the lens of ion channels.

SEC31 homolog A (SEC31A) is associated with a variety of cancers. This study elucidated the role of SEC31A in gastric cancer (GC). The expression of SEC31A in GC was analyzed using western blotting and immunohistochemistry. Functional experiments were performed to detect the impact of SEC31A on GC proliferation, migration and invasion. The PI3K inhibitor LY294002 was used to explore the regulation of the biological functions by SEC31A through the PI3K/AKT/mTOR pathway in GC cells. Co‑culture of THP‑1 (human leukemia cell line of monocytic origin) and GC cells was conducted to investigate the impact of SEC31A on the M2 macrophage polarization in tumor immune microenvironment. Our investigation observed elevated expression of SEC31A in both GC tissue and cell lines. Through functional studies, suppression of SEC31A was found to decrease epithelial‑mesenchymal transition (EMT), proliferation, migration and invasion in GC cells. Overexpression of SEC31A led to activation of the PI3K/AKT/mTOR signaling cascade. The PI3K inhibitor LY294002 counteracted the increases in proliferation, migration, invasion and EMT triggered by SEC31A. Co‑culture experiments involving THP‑1 and GC cells indicated that SEC31A enhanced the recruitment and M2 polarization of macrophages via CCL5 within the GC tumor microenvironment (ME). To assess the influence of M2 macrophages on malignant behavior of GC cells, M2 macrophage‑conditioned medium (M2‑CM) was applied. M2‑CM mitigated the inhibitory effects on malignant phenotypes in GC cells following SEC31A knockdown. Moreover, diminished SEC31A expression enhanced the efficacy of anti‑PD‑1 therapy in GC models. Collectively, SEC31A promoted GC progression via activating the PI3K/AKT/mTOR pathway and facilitating CCL5‑mediated M2 polarization and recruitment of tumor‑associated macrophages. SEC31A knockdown may augment the anti‑tumor response to anti‑PD‑1 treatment in GC. These findings highlight SEC31A as a valuable prognostic indicator and potential target for immunotherapy in GC, offering novel strategies to enhance the effectiveness of anti‑PD‑1 therapy.