Background/Aims: Dysregulated cholesterol metabolism is a hallmark of hepatocellular carcinoma (HCC) that drives tumor initiation and progression. However, clinical targeting of cholesterol metabolism has yielded limited benefits due to stringent feedback in tumor cells. Identifying a central mediator capable of restoring cholesterol homeostasis within the cell’s intrinsically fine-tuned regulatory framework is urgently needed. Methods: We integrated a proteomic dataset from patients with cholesterol-dysregulated HCC into a global cholesterol metabolic regulatory network to identify potential therapeutic targets for disrupted cholesterol homeostasis. The prognostic significance of the candidate targets was further validated in an independent cohort through immunohistochemistry. Functional and mechanistic studies were conducted in vitro using HCC cell lines and in vivo using mouse models. The pharmacological efficacy of the candidate agent was evaluated in both subcutaneous and orthotopic HCC mouse models. Results: ER lipid raft-associated 1 (ERLIN1), a pivotal regulator of cholesterol metabolism reprogramming, was identified as an independent favorable prognostic indicator in HCC. ERLIN1 constrains HCC progression both in vitro and in vivo by stabilizing the INSIG1–SCAP–SREBP2 axis and maintaining the metabolic balance of intracellular cholesterol. Under hypoxia, impaired factor-inhibiting hypoxia-1-dependent hydroxylation of ASB11 at asparagine residues 90 and 92 enhances ASB11-mediated ERLIN1 degradation. Pharmacological targeting of this axis using zoledronic acid (ZoA) attenuated HCC progression by weakening the ASB11–ERLIN1 interaction and restoring cholesterol homeostasis. Conclusions ERLIN1 represents a druggable metabolic vulnerability in cholesterol-dysregulated HCC. Targeting the ASB11–ERLIN1 axis with the clinically approved ZoA reestablishes cholesterol homeostasis and offers a promising therapeutic strategy to overcome the current limitations of cholesterol-targeted HCC therapies.

Objective To explore the mechanism by which the early growth response protein 1(EGR1)/AMP-activated protein kinase(AMPK)/nuclear factor erythroid 2-related factor 2(Nrf2)pathway improves Neuro-2a cell injury via Mendelian randomization(MR),bioinformatics and in vitro models of ischemic stroke(IS).Methods The blood proteins associated with genetic susceptibility to IS were identified based on MR analysis.Then,the GSE22255 dataset was comprehensively analyzed,including GSEA,immunoinfiltration and differential expression analysis.The transcription factors closely related to the occurrence and development of IS were identified after joint analysis with MR results,and the biological functions of key genes were further verified by in vitro experiments.Results A total of 712 proteins related to IS susceptibility were identified by MR analysis.A total of 2357 differentially expressed genes were identified from the GSE22255 dataset,and 75 intersection genes and 34 transcription factors were identified after combined analysis with MR results.In vitro experiments showed that knockdown of EGR1 expression could significantly inhibit oxygen-glucose deprivation/reperfusion(OGD/R)-induced Neuro-2a cell damage,and the mechanism might be related to the up-regulation of p-AMPK and Nrf2 protein expression.Conclusion This study integrates MR analysis,transcriptomics and in vitro experiments to reveal the potential role of 75 IS susceptibility genes and transcription factor EGR1 in the pathogenesis of IS,which provides theoretical basis for the development of therapeutic drugs for IS.

Decreased endometrial receptivity is a critical factor contributing to the decline in fertility among women of advanced age. With the increase of age, the endometrium exhibits diminished hormonal responsiveness, a phenomenon known as inflammaging, and an imbalance in immune cell populations. These changes impede the processes of endometrial epithelial-mesenchymal transition and decidualization, thereby negatively impacting embryo implantation and reducing overall fertility potential. Furthermore, adverse lifestyle choices, exposure to environmental factors, inflammatory conditions, and repeated interventions within the uterine cavity can result in pathological aging of the endometrium that does not align with its chronological age. This misalignment may be associated with the emergence of various reproductive disorders, even in younger women. This paper seeks to investigate the relationship between pathological aging of the endometrium and reproductive disorders, particularly in the context of embryo implantation, with the aim of providing new insights into the pathogenesis, clinical diagnosis, and treatment of reproductive disorders.

Objective:To evaluate the therapeutic efficacy of the Wenyang Huazhuo Formula on spontaneous pregnancy outcomes in advanced-age infertile patients with spleen-kidney yang deficiency syndrome.Methods:A retrospective cohort study design was employed to gather clinical data from 303 advanced-age infertile patients with spleen-kidney yang deficiency syndrome who sought treatment at the Department of Reproductive Medicine in the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine between June 2019 and June 2023. Patients were categorized into two groups based on their treatment regiments: a basic treatment group (non-exposed group, n=151) and a group receiving basic treatment in conjunction with the Wenyang Huazhuo Formula (exposed group, n=152). The study compared and analyzed pregnancy outcomes, including time to pregnancy (TTP) within one year, ongoing pregnancy rate, and clinical pregnancy rate, between the two groups over a one-year follow-up period. Additionally, stratified analyses were performed based on age and anti-Müllerian hormone (AMH) levels to evaluate the impact of Wenyang Huazhuo Formula on pregnancy outcomes across different patient subgroups. Results:The TTP in the exposed group was significantly reduced compared with the non-exposed group ( HR=1.70, 95% CI: 1.12-2.57, P=0.013), suggesting a significant shortening of their TTP, and the ongoing pregnancy rate [28.29% (43/152)] and the clinical pregnancy rate [33.55% (51/152)] were significantly higher than those in the non-exposed group [13.91% (21/151), P=0.020; 21.85% (33/151), P=0.023], and the early abortion rate [15.69% (8/51)] was significantly reduced compared with the non-exposed group [36.36% (12/33), P=0.031]. Stratified analyses by age revealed that for patients aged 35-37 years, the exposed group exhibited a significantly shorter TTP ( HR=1.67, 95% CI: 1.03-2.70, P=0.038), while no significant difference in TTP was observed in the 38-44-year old patients ( HR=1.78, 95% CI: 0.77-4.10, P=0.174). Regarding the ongoing pregnancy rate, the exposed group showed a significant higher rate [31.31% (31/99)] than the non-exposed group [13.91% (15/112), P=0.022] among patients aged 35-37 years, whereas no significant difference was observed in the 38-44 years old group ( P=0.386). Stratified analysis based on AMH levels showed that both low AMH levels (<1.68 μg/L) and high AMH levels (≥1.68 μg/L) patients in the exposed group experienced a significant reduction in TTP compared with the non-exposed group ( HR=8.07, 95% CI: 1.04-62.52, P=0.046; HR=1.57, 95% CI: 1.01-2.43, P=0.044, respectively), along with a notable improvement in the ongoing pregnancy rate [13.64% (6/44) vs. 0% (0/29), P=0.038; 34.26% (37/108) vs.17.21% (21/122), P=0.028, respectively]. Further stratified analysis indicated that, compared with patients with low AMH levels, both the exposed group and the non-exposed group of patients with high AMH levels exhibited a significantly shorter TTP ( HR=1.91, 95% CI: 1.02-3.69, P=0.043; HR=9.48, 95% CI: 1.30-69.22, P=0.002, respectively), along with a significantly higher ongoing pregnancy rate ( P=0.014; P=0.008). Conclusion:The Wenyang Huazhuo Formula significantly shortens TTP, enhances both the clinical pregnancy rate and the ongoing pregnancy rate and reduces the early abortion rate in advanced-age infertile women diagnosed with spleen-kidney yang deficiency syndrome.

Objective:To systematically assess qualitative studies exploring the experiences of postpartum lifestyle changes among women with hypertensive disorders of pregnancy, comprehensively integrate these experiences, and provide a foundational reference for the subsequent development of targeted healthy lifestyle interventions.Methods:A comprehensive computerized search was conducted across PubMed, Embase, Web of Science, Cochrane Library, Pro quest, CINAHL, China National Knowledge Infrastructure, Wanfang Database, VIP, and China Biomedical Database, encompassing all qualitative studies that met the inclusion criteria from the inception of the databases to June 21st 2024. The quality of the retrieved literature was evaluated using the Centre for Evidence-Based Health Care Quality Assessment Criteria for Qualitative Research at the Joanna Briggs Institute, Australia. The results were integrated using the pooled integration method, and the quality of the integrated findings was rated using the ConQual method.Results:A total of 10 papers were included, and 23 findings were distilled and grouped into 9 new categories, resulting in 4 integrative findings: multiple dilemmas and challenges to postpartum lifestyle change, postpartum lifestyle change with the change of mother′s role, facilitators of postpartum lifestyle change, and impediments to postpartum lifestyle change.Conclusions:Postnatal lifestyle change in women with hypertensive disorders of pregnancy is crucial for improving prognosis. In the future, healthcare professionals should improve the social support system, promote postnatal role change to adapt to the new life, improve women′s awareness of risk and disease, maintain the continuity of postnatal follow-up, and incorporate a variety of behavioural change strategies to stimulate the motivation for lifestyle change, so as to promote the effective change of postnatal lifestyles.

Objective:To investigate and analyze the current status of dietary behavior compliance among patients following bariatric metabolic surgery, as well as its associated influencing factors.Methods:A convenience sampling approach was employed to select 149 discharged patients who underwent bariatric metabolic surgery at the Gastric Surgery Department of a tertiary hospital in Fuzhou between August 2021 and February 2024. A WeChat communication group was established for survey participants, and a questionnaire survey was conducted via the mobile app "Wenjuanxing." Patients′ data were collected using the General Information Questionnaire and the Dietary Behavior Compliance Scale for Patients After Bariatric Metabolic Surgery. Multiple stepwise linear regression analysis was used to evaluate the influencing factors of dietary behavior compliance in postoperative patients.Results:A total of 165 questionnaires were distributed, with 149 valid responses retrieved. Among the 149 post-bariatric surgery patients, 37 were males and 112 were females, with an average age of (31.10 ± 7.26) years. The overall level of dietary behavior compliance was (56.91 ± 9.70). Significant differences in dietary behavior compliance were observed among patients of different genders, educational levels, drinking habits, and postoperative durations ( t = 2.17, 1.93, F = 4.27, 3.50, all P<0.05). Specifically, postoperative duration emerged as the primary factor influencing dietary behavior compliance following bariatric metabolic surgery ( t = -2.22, P<0.05). Conclusions:Female patients, those with a college degree or higher, and individuals within 0 to 6 months post-surgery demonstrated better dietary behavior compliance. Medical staff should comprehensively consider patient individual differences, develop, and implement personalized health education plans to enhance patient dietary behavior compliance.

The amino acid sequences of the OmpA protein isolated from Escherichia coli QML2206-1(E.coli QML2206-1)in our laboratory were analyzed for homology with different strains of OmpA proteins using bioinformatics software,and the OmpA protein was analyzed for its physicochemical properties,transmembrane structure and signal peptide prediction,B-cell anti-genic epitope prediction,secondary and tertiary structure prediction.The OmpA gene fragment was ligated with pET-32a vector to construct a prokaryotic expression vector,which was purified by a nickel column affinity purification system after prokaryotic expression and optimization of ex-pression conditions in BL21(DE3).The purified recombinant protein was fully mixed with Freund's adjuvant to immunize mice,and the levels of mouse-specific IgG antibody and the expression levels of cytokines CD4,CD8 and IL-4 in mouse serum were detected by ELISA,and the immuno-protective effect was evaluated by mouse attack protection test.OmpA protein is a hydrophilic protein with no transmembrane structural domains and a secondary structure consisting mainly of irregular coils(47.98％)and α-helices(29.77％),with 12 antigenic epitopes that can bind to anti-bodies produced by B cells.The recombinant protein OmpA with a relative molecular mass of a-bout 55 kDa was successfully obtained by prokaryotic expression,and the highest expression was induced by IPTG concentration of 0.000 4 mmol/L for 6 h at 37 ℃.The serum-specific IgG anti-body potency of recombinant protein immunized mice was up to 1∶32 000;the expression levels of CD4,CD8 and IL-4 in the serum of immunized mice were elevated compared with those of the con-trol group.The survival rate of mice was 80％and 40％after attack with minimum lethal dose(MLD)and 2 times minimum lethal dose(2MLD),respectively.OmpA recombinant protein has good antigenicity and certain immunoprotective effects,and this study provides a technical basis for the next step in the development of a genetically engineered subunit vaccine against yak-appli-cable E.coli based on OmpA protein.

OBJECTIVE To explore the clinical characteristics of Tropheryma whipplei(TW)infection and observe the application of pathogenic metagenomic next-generation sequencing(mNGS)in diagnosis of TW infection.METHODS The clinical data were collected from 1 patient who was diagnosed by mNGS in the Affiliated Hospital of Jining Medical University on Apr.9,2022.The data including the results of laboratory tests and treatment out-comes were summarized,and a literature review was conducted.RESULTS A 50-year-old woman presented to the hospital with chest tightness and chest pain lasting for 3 days,accompanied by dyspnea,palpitations,and expec-toration.The chest plain CT scan and magnetic resonance imaging(MRI)scan suggested a high probability of pul-monary infection.Normal flora were isolated by culture of bronchoalveolar lavage fluid(BALF);TW and human βherpes virus type 7 were detected in BALF by mNGS,with the sequence numbers 327 000 and 9,respectively.The pulmonary symptoms of the patient were improved after joint treatment of the infection with etimicin,levo-floxacin and minocycline.The patient repeatedly sought for medical treatment due to the pain of shoulder joint and limitation of motion.CONCLUSIONS TW is one of major pathogens leading to the infections of systemic multiple systems,and it is necessary to attach great importance to the diagnosis and treatment.The traditional laboratory test method can not achieve ideal diagnosis effect and is more likely to make a missed diagnosis.mNGS is more ac-curate and more efficient than the traditional detection method in diagnosis of TW-induced diseases.Early use of mNGS can make a rapid identification of pathogens and facilitate the reasonable clinical use of antibiotics.It is of great significance for control of the disease progression,improvement of prognosis and prevention of recurrence.

Objective:To evaluate the therapeutic efficacy of the Wenyang Huazhuo Formula on spontaneous pregnancy outcomes in advanced-age infertile patients with spleen-kidney yang deficiency syndrome.Methods:A retrospective cohort study design was employed to gather clinical data from 303 advanced-age infertile patients with spleen-kidney yang deficiency syndrome who sought treatment at the Department of Reproductive Medicine in the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine between June 2019 and June 2023. Patients were categorized into two groups based on their treatment regiments: a basic treatment group (non-exposed group, n=151) and a group receiving basic treatment in conjunction with the Wenyang Huazhuo Formula (exposed group, n=152). The study compared and analyzed pregnancy outcomes, including time to pregnancy (TTP) within one year, ongoing pregnancy rate, and clinical pregnancy rate, between the two groups over a one-year follow-up period. Additionally, stratified analyses were performed based on age and anti-Müllerian hormone (AMH) levels to evaluate the impact of Wenyang Huazhuo Formula on pregnancy outcomes across different patient subgroups. Results:The TTP in the exposed group was significantly reduced compared with the non-exposed group ( HR=1.70, 95% CI: 1.12-2.57, P=0.013), suggesting a significant shortening of their TTP, and the ongoing pregnancy rate [28.29% (43/152)] and the clinical pregnancy rate [33.55% (51/152)] were significantly higher than those in the non-exposed group [13.91% (21/151), P=0.020; 21.85% (33/151), P=0.023], and the early abortion rate [15.69% (8/51)] was significantly reduced compared with the non-exposed group [36.36% (12/33), P=0.031]. Stratified analyses by age revealed that for patients aged 35-37 years, the exposed group exhibited a significantly shorter TTP ( HR=1.67, 95% CI: 1.03-2.70, P=0.038), while no significant difference in TTP was observed in the 38-44-year old patients ( HR=1.78, 95% CI: 0.77-4.10, P=0.174). Regarding the ongoing pregnancy rate, the exposed group showed a significant higher rate [31.31% (31/99)] than the non-exposed group [13.91% (15/112), P=0.022] among patients aged 35-37 years, whereas no significant difference was observed in the 38-44 years old group ( P=0.386). Stratified analysis based on AMH levels showed that both low AMH levels (<1.68 μg/L) and high AMH levels (≥1.68 μg/L) patients in the exposed group experienced a significant reduction in TTP compared with the non-exposed group ( HR=8.07, 95% CI: 1.04-62.52, P=0.046; HR=1.57, 95% CI: 1.01-2.43, P=0.044, respectively), along with a notable improvement in the ongoing pregnancy rate [13.64% (6/44) vs. 0% (0/29), P=0.038; 34.26% (37/108) vs.17.21% (21/122), P=0.028, respectively]. Further stratified analysis indicated that, compared with patients with low AMH levels, both the exposed group and the non-exposed group of patients with high AMH levels exhibited a significantly shorter TTP ( HR=1.91, 95% CI: 1.02-3.69, P=0.043; HR=9.48, 95% CI: 1.30-69.22, P=0.002, respectively), along with a significantly higher ongoing pregnancy rate ( P=0.014; P=0.008). Conclusion:The Wenyang Huazhuo Formula significantly shortens TTP, enhances both the clinical pregnancy rate and the ongoing pregnancy rate and reduces the early abortion rate in advanced-age infertile women diagnosed with spleen-kidney yang deficiency syndrome.

ObjectiveTo explore the possible mechanisms of Wenyang Huazhuo Formula （温阳化浊方， WHF） in improving reproductive aging from the perspective of the ovarian mechanical microenvironment. MethodsThe experiment included five groups， 3-month group （20 female mice at 3 months of age）， 6-month group （20 female mice at 6 months of age）， 6-month + WHF group （20 female mice at 5 months of age treated with WHF）， 9-month group （20 female mice at 9 months of age）， and 9-month + WHF group （20 female mice at 8 months of age treated with WHF）. The 6-month + WHF group and 9-month + WHF group were orally administered WHF 41.2 g/（kg·d） once daily for 4 consecutive weeks. The other three groups received no intervention. Reproductive hormone levels were measured by ELISA. HE staining was used to count the numbers of various stages of follicles. Ovarian hyaluronic acid （HA） content and collagen fiber content were measured to evaluate the ovarian mechanical microenvironment. Superovulation was performed to observe the number of eggs obtained， as well as the number of offspring and birth weight to assess fertility. The in vitro fertilization and blastocyst culture of oocytes from female offspring in each group were observed to evaluate the effect of WHF on offspring reproductive potential. ResultsCompared with the 3-month group， the 6-month group and 9-month group showed significantly decreased serum levels of gonadotropin-releasing hormone （GnRH）， follicle-stimulating hormone （FSH）， and luteinizing hormone （LH）， decreased ovarian collagen content， and reduced numbers of primordial and secondary follicles. In contrast， the numbers of primary follicles， antral follicles， and atretic follicles increased. The levels of anti-Müllerian hormone （AMH）， ovarian HA content， and the fertilization rate， cleavage rate， and blastocyst formation rate of oocytes from offspring were significantly lower （P＜0.05）. Compared with the 6-month group， the 6-month + WHF group showed significantly reduced serum levels of GnRH， FSH， and LH， with a significant decrease in primary follicles， antral follicles， and atretic follicles as well as increase of AMH levels， ovarian HA content， number of primordial and secondary follicle， egg count， and offspring birth weight （P＜0.05）. Compared with the 9-month group， the 9-month + WHF group exhibited reduced GnRH， FSH， and collagen fiber content， as well as reduced number of primary follicles， antral follicles， and atretic follicles. However， AMH levels， ovarian HA content， number of primordial and secondary follicle， egg count， offspring numbers， birth weight， fertilization rate， cleavage rate， and blastocyst formation rate of oocytes from offspring all significantly increased （P＜0.05）. ConclusionWHF can significantly improve the ovarian reserve， fertility， and reproductive potential in offspring during reproductive mid-life and late-life stages. Its effect may be related to the remodeling of the mechanical microenvironment of aging ovaries. Moreover， the effect on the mechanical microenvironment remodeling of late-stage ovaries and the improvement of the offspring reproductive potential is more significant.