eIF3a is a N ⁶-methyladenosine (m⁶A) reader that regulates mRNA translation by recognizing m⁶A modifications of these mRNAs. It has been suggested that eIF3a may play an important role in regulating translation initiation via m⁶A during infection when canonical cap-dependent initiation is inhibited. However, the death of animal model studies impedes our understanding of the functional significance of eIF3a in immunity and regulation in vivo. In this study, we investigated the in vivo function of eIF3a using eIF3a knockout and knockdown mouse models and found that eIF3a deficiency resulted in splenic tissue structural disruption and multi-organ damage, which contributed to severe sepsis induced by Lipopolysaccharide (LPS). Ectopic eIF3a overexpression in the eIF3a knockdown mice rescued mice from LPS-induced severe sepsis. We further showed that eIF3a maintains a functional and healthy immune system by regulating B cell function and quantity through m⁶A modification of mRNAs. These findings unveil a novel mechanism underlying sepsis, implicating the pivotal role of B cells in this complex disease process regulated by eIF3a. Furthermore, eIF3a may be used to develop a potential strategy for treating sepsis.

AIM:To explore the protective effect of total flavonoids from rhododendron(TFR)on oxi-dative stress injury in the brain of rats subjected to cerebral ischemia/reperfusion(I/R).METHODS:For-ty male SD rats were randomly divided into 5 groups:sham operation group(Sham group),cere-bral ischemia-reperfusion group(MCAO group),and Cerebral Ischemia/Reperfusion with TFR treat-ment groups(TFR 50,100,200 mg/kg groups).The MCAO group and TFR-treated groups underwent ischemia/reperfusion surgery,and the TFR-treated groups received TFR intragastrically for 14 consecu-tive days following the ischemia/reperfusion injury.After 14 days,comparisons were made in terms of neurological function scores,serum inflammatory factors,oxidative stress indicators,and brain injury markers.Additionally,histological examination of brain tissue morphology using Hematoxylin and Eo-sin(HE)staining,observation of cerebral blood flow through cerebral blood flow imaging,and measurement of lactate dehydrogenase(LDH),neu-ron-specific enolase(NSE)activity in serum,inter-leukin-1(IL-1),interleukin-6(IL-6)levels,superox-ide dismutase(SOD)activity,malondialdehyde(MDA)content,nitric oxide(NO)content,and nitric oxide synthase(NOS)activity were performed.RE-SULTS:Compared to the Sham group,MCAO rats exhibited abnormal neurological function scores,severe damage to the microstructure of brain tis-sue,noticeable changes in brain morphology,ele-vated activities of LDH and NSE,increased levels of IL-1 and IL-6,elevated MDA content,and de-creased SOD,NOS activity,and NO content.In com-parison to the MCAO group,rats treated with TFR at doses of 50,100,and 200 mg/kg showed recov-ery of abnormal neurological function scores,re-duced damage to the microstructure of brain tis-sue,decreased activities of LDH and NSE,lowered levels of IL-1 and IL-6,reduced MDA content,and increased SOD,NOS activity,and NO content.CON-CLUSION:Total flavonoids from Rhododendron can protect against cerebral ischemia/reperfusion inju-ry,reducing oxidative stress levels.

Objective:To explore the correlation between serum uric acid/high-density lipoprotein cholesterol ratio (UHR) and the risk of hypertension in elderly physical examination populations.Methods:This study was a cross-sectional study. A total of 1 028 patients aged≥60 years who underwent physical examinations at the Health Management Center of Fuwai Central China Cardiovascular Hospital from September 2023 to February 2024 were included in this study. The general demographic data, past medical history, physical examination and laboratory examination indicators of the physical examiners were collected, and according to whether they had hypertension or not, they were divided into hypertension group (390 cases) and non-hypertension group (638 cases), and all UHR values were arranged from small to large, and the UHR was divided into three groups by tertiles of UHR, and the general data and blood biochemical indexes between the groups were compared. Spearman rank correlation was used to analyze the correlation between UHR level and body mass index, total cholesterol, triglyceride and other indexes in the elderly population. Logistic regression was used to explore the relationship between UHR level and hypertension in the elderly population, and the stratification analysis of the physical examination population was carried out according to diabetes, coronary heart disease and dyslipidemia, and the interaction test between groups was carried out.Results:Among the 1 028 geriatric physical examination cases, 580 (56.4%) were males and 448 (43.6%) were females, aged (66.7±5.8) years. UHR levels were higher in the hypertensive group compared to the non-hypertensive group [248.88 (191.19, 322.25) vs 213.52 (165.94, 275.29); Z=-5.445, P<0.05]. With the increase of UHR level, the detection rate of hypertension in the elderly population increased (accounted for 27.8%, 38.2% and 47.8%, respectively; χ2=29.211, P<0.05). Spearman rank correlation analysis showed that UHR was positively correlated with body mass index, triglycerides, serum uric acid, serum creatinine and fasting blood glucose ( r=0.318, 0.334, 0.774, 0.474, 0.080; all P<0.05), and negatively correlated with total cholesterol, glomerular filtration rate and low-density lipoprotein cholesterol ( r=-0.239, -0.303, -0.154; all P<0.05). When the confounding factors were not adjusted (model 1), the risk of hypertension in high UHR group was 2.382 times higher than that in low UHR group and 1.607 times higher than that in medium UHR group; after adjusting for all confounding factors such as age, gender, body mass index, systolic blood pressure, diastolic blood pressure, junior high school education or below, smoking, alcohol consumption, glomerular filtration rate, etc., the risk of hypertension in the high-level UHR group was 1.732 times higher than that in the low-level UHR group (95% CI: 1.139-2.635) ( P<0.05). The elderly physical examination population was further stratified according to whether there was diabetes, dyslipidemia and coronary heart disease, and it was found that there was no interaction between UHR and diabetes, dyslipidemia and coronary heart disease on the prevalence of hypertension (all P>0.05). Conclusions:Hypertension detection rate increases with higher UHR levels. UHR is closely related to the incidence of hypertension in the elderly population.

Trophoblast cells serve as the foundation for placental development. We analyzed published multiomics sequencing data and found that trophoblast cells highly expressed RRS1 compared to primitive endoderm and epiblast. We used HTR-8/SVneo cells for further investigation, and Western blot and immunofluorescence staining confirmed that HTR-8/SVneo cells highly expressed RRS1. RRS1 was successfully knocked down in HTR-8/SVneo cells using siRNA. Using IncuCyte S3 live-cell analysis system based on continuous live-cell imaging and real-time data, we observed that proliferation, migration, and invasion abilities were all significantly decreased in RRS1-knockdown cells. RNA-seq revealed that knockdown of RRS1 affected the gene transcription, and upregulated pathways in extracellular matrix organization, DNA damage response, and intrinsic apoptotic signaling, downregulated pathways in embryo implantation, trophoblast cell migration, and wound healing. Differentially expressed genes were enriched in diseases related to placental development. Consistent with these findings, human chorionic villus samples collected from spontaneous abortion cases exhibited significantly reduced RRS1 expression compared to normal controls. Our results highlight the functional importance of RRS1 in human trophoblasts and suggest that its deficiency contributes to early pregnancy loss.
Humans ; Trophoblasts/physiology* ; Cell Movement/genetics* ; Cell Proliferation/genetics* ; Female ; Pregnancy ; Abortion, Spontaneous/metabolism* ; Cell Line ; Placentation/genetics*

Abstract Alternative splicing(AS) is one of the important ways of post-transcriptional regulation, which can generate multiple mRNA isoforms from a single gene. In recent years, many studies have shown that AS can occur in almost all types of tumors, and the abnormal expression of splicing factors is related to the disease progression of tumor patients, which may become a potential marker for judging tumor progression. Therefore, this review summarizes the role, molecular mechanism, clinical relevance and treatment response of AS in tumors. It is found that AS can participate in the progression of malignant tumors by regulating tumor invasion, metastasis, apoptosis, cell metabolism, and promoting tumor immune escape and treatment resistance, which provides a theoretical basis for the clinical application of AS. The precise identification of tumor-specific AS and the development of small molecule inhibitors targeting specific AS isoforms may provide a new direction for precise and personalized cancer treatment.

Objective : To explore the role of semaphoring 6d(SEMA6D) in the malignant progression of triple-negative breast cancer(TNBC). Methods : Bioinformatics and Immunohistochemistry(IHC) were used to analyze the expression level of SEMA6D in TNBC and paracancer non-tumor tissues and its relationship with patients′ clinicopathological features. MDA-MB-231 cell line stably knocking down the expression of SEMA6D was constructed, and the effects of SEMA6D on migration and invasion of TNBC cells were investigated by Wound-healing assays and Transwell assays. cBioPortal and GEPIA2 databases were used to screen out the gene negatively associated with it, namely aurora kinase A(AURKA). Bioinformatics and IHC were used to analyze the expression level of AURKA in TNBC and paracancer non-tumor tissues and its relationship with patients' clinicopathological features. Western blot assay was used to analyze the expression of AURKA and the effect of epithelial-mesenchymal transition(EMT) makers Claudin-1, N-cadherin and Vimentin after knocking downSEMA6D. Results: Bioinformatics analysis and IHC results showed that the expression of SEMA6D in TNBC tissues was significantly lower than that in paracancer non-tumor tissues(bothP<0.05). The expression of AURKA in TNBC tissues was significantly higher than that in paracancer non-tumor tissues(bothP<0.05), SEMA6D and AURKA were significantly negatively correlated in TNBC(P<0.01). Both low expression of SEMA6D and high expression of AURKA were positively correlated with tumor size, tumor histological grade, clinical stage and lymph node metastasis in TNBC patients(allP<0.05). The knockdown ofSEMA6Dsignificantly promoted the migration and invasion ability of TNBC cells(bothP<0.01). Western blot results showed that the knockdown ofSEMA6Dupregulated AURKA expression, promoted the expression of N-cadherin and Vimentin, and inhibited the expression of Claudin-1 in tumor cells. Conclusion Down-regulation of SEMA6D expression in TNBC may be involved in the malignant progression of TNBC through up-regulation of AURKA expression and promotion of EMT.

Objective : To explore the expression of prohibitin2(PHB2) in breast cancer and its effect on the biological behaviors of tumor cells. Methods : Immunohistochemistry was used to detect the expression of PHB2 protein in breast cancer tissues and its relationship with clinicopathologic features. Breast cancer stable transient cell lines were constructed with knockdown and overexpression ofPHB2, respectively. The effects of PHB2 on cell proliferation, migration and invasion ability were detected by clone formation assay, scratch assay and Transwell assay. Western blot(WB) was used to detect the effects of PHB2 on the expression of epithelial-mesenchymal transition(EMT)-related markers, including E-cadherin, N-cadherin, Snail family transcriptional repressor 1(Snail) protein, Vimentin, and Claudin-1. The effect of PHB2 on tumorigenicityin vivowas detected by subcutaneous tumor formation assay in nude mice. Results: The result of immunohistochemical showed that PHB2 was highly expressed in breast cancer and the expression of PHB2 was significantly positive correlated with tumor size, human epidermal growth factor receptor-2(HER-2) status and proliferation index Ki-67 levels(P<0.05). Clone formation assay, scratch assay and Transwell assay revealed that knockdown ofPBH2significantly inhibited the proliferation, migration and invasion ability of breast cancer cells(P<0.01), while the overexpression ofPHB2significantly promoted cell proliferation, migration and invasion(P<0.01). The result of subcutaneous tumor formation experiment in nude mice revealed a significant decrease in tumor volume and weight in knockdownPHB2mice(P<0.000 1), whilePHB2overexpression tumors significantly increased in volume and weight(P<0.001).WB assay showed that the protein expression of epithelial marker E-cadherin increased, while the expressions of mesenchymal markers N-cadherin, Snail and Vimentin decreased significantly afterPHB2knockdown with them in control cells(P<0.01). The expression of Claudin-1 decreased, while the expressions of N-cadherin, Snail and Vimentin increased significantly inPHB2overexpression cells(P<0.05). Conclusion PHB2 is highly expressed in breast cancer and promotes multiple malignant biological behaviors in tumor cells, suggesting PHB2 may be a potential target for breast cancer diagnosis and treatment.

Objective : To investigate the expression of Keratin 14(KRT14) in Basal-like Breast Cancer(BLBC) and its biological functions and mechanisms. Methods : The expression levels of KRT14 mRNA in BLBC and para-cancer breast tissues were analyzed using The Cancer Genome Atlas(TCGA) database. qPCR, Western blot(WB), and immunohistochemistry were employed to detect KRT14 expression in BLBC and adjacent normal tissues, and its correlation with clinicopathological features was analyzed. KRT14 overexpression and knockdown were performed in breast cancer cells, and cell scratch and transwell assays were performed to evaluate changes in migration and invasion abilities. To investigate the expression of proteins related to the Wnt/β-catenin signaling pathway, including catenin Beta 1(β-catenin), wingless-type MMTV integration site family, member 1(Wnt1), matrix metallopeptidase 7(MMP7), and cellular myelocytomatosis viral oncogene homolog(c-Myc), as well as the cellular localization of β-catenin, WB and immunofluorescence(IF) techniques were employed. Additionally, a Wnt/β-catenin signaling pathway inhibitor was used to verify the mechanism of action of KRT14. Results : The expression of KRT14 was significantly higher in BLBC tissues compared to normal tissues(P<0.05), and was associated with higher T stage and histological grade(P<0.05). The overexpression of KRT14 significantly enhanced the migration and invasion abilities of breast cancer cells, while the knockdown of KRT14 significantly reduced those abilities(P<0.01). The overexpression of KRT14 can increase the expression levels of Wnt/β-catenin pathway-related proteins β-catenin, Wnt1, MMP7, and c-Myc, thereby activating the Wnt/β-catenin pathway. Moreover, the inhibition of this pathway can eliminate the effects of KRT14 on cell migration and invasion. Conclusion The high expression of KRT14 in BLBC may promote the migration and invasion of breast cancer cells through the Wnt/β-catenin signaling pathway.

Objective : To investigate the expression of(heat shock protein a member 8,HSPA8) in breast cancer and its effect on tumor biological behaviors. Methods: Bioinformatics analysis and immunohistochemistry assays were used to detect the expression of HSPA8 in breast cancer and adjacent non-tumor breast tissues,and the relationship between its expression and clinicopathological characteristics of breast cancer patients was analyzed.Correlation between HSPA8 expression and prognosis of breast cancer patients was analyzed by Kaplan-Meier Plotter database.HSPA8 knockdown and over expression breast cancer stabilized cells were constructed,respectively.CCK-8,clone formation,Transwell,cell scratch,Western blot and immunofluorescence assay were used to detect the effects of HSPA8 on the proliferation,invasion and migration of breast cancer cells,and its effect on epithelial-mesenchymal transition(EMT). Results : Bioinformatics analysis and immunohistochemistry assay revealed that the expression of HSPA8 in breast cancer tissues was higher than that in adjacent non-tumour tissues(P<0.05),and its expression level of the protein was significantly and positively correlated with the tumor size,histological grade,lymph node metastasis and Ki-67 proliferation index(P<0.05).The Kaplan-Meier survival curve showed that high expression of HSPA8 was significantly associated with poor prognosis in breast cancer patients(P<0.05).CCK-8,clone formation,transwell,cell scratch,Western blot and immunofluorescence assay showed that knockdown of HSPA8 expression could significantly inhibit the proliferation,invasion,migration function and EMT of breast cancer cells(P<0.05),while overexpression of HSPA8 could significantly promote the proliferation,invasion,migration function and EMT of breast cancer cells(P<0.05). Conclusion HSPA8 is highly expressed in breast cancer tissues,which is closely related to disease progression and the malignant phenotype of breast cancer,suggesting that HSPA8 may be a potential biological target for breast cancer treatment.

AIM:To explore the protective effect of total flavonoids from rhododendron(TFR)on oxi-dative stress injury in the brain of rats subjected to cerebral ischemia/reperfusion(I/R).METHODS:For-ty male SD rats were randomly divided into 5 groups:sham operation group(Sham group),cere-bral ischemia-reperfusion group(MCAO group),and Cerebral Ischemia/Reperfusion with TFR treat-ment groups(TFR 50,100,200 mg/kg groups).The MCAO group and TFR-treated groups underwent ischemia/reperfusion surgery,and the TFR-treated groups received TFR intragastrically for 14 consecu-tive days following the ischemia/reperfusion injury.After 14 days,comparisons were made in terms of neurological function scores,serum inflammatory factors,oxidative stress indicators,and brain injury markers.Additionally,histological examination of brain tissue morphology using Hematoxylin and Eo-sin(HE)staining,observation of cerebral blood flow through cerebral blood flow imaging,and measurement of lactate dehydrogenase(LDH),neu-ron-specific enolase(NSE)activity in serum,inter-leukin-1(IL-1),interleukin-6(IL-6)levels,superox-ide dismutase(SOD)activity,malondialdehyde(MDA)content,nitric oxide(NO)content,and nitric oxide synthase(NOS)activity were performed.RE-SULTS:Compared to the Sham group,MCAO rats exhibited abnormal neurological function scores,severe damage to the microstructure of brain tis-sue,noticeable changes in brain morphology,ele-vated activities of LDH and NSE,increased levels of IL-1 and IL-6,elevated MDA content,and de-creased SOD,NOS activity,and NO content.In com-parison to the MCAO group,rats treated with TFR at doses of 50,100,and 200 mg/kg showed recov-ery of abnormal neurological function scores,re-duced damage to the microstructure of brain tis-sue,decreased activities of LDH and NSE,lowered levels of IL-1 and IL-6,reduced MDA content,and increased SOD,NOS activity,and NO content.CON-CLUSION:Total flavonoids from Rhododendron can protect against cerebral ischemia/reperfusion inju-ry,reducing oxidative stress levels.