Min Zhang

Introduction: Intrahepatic cholestasis of pregnancy (ICP) is a pregnancy-specific liver disorder. Serum total bile acids (TBA) are the primary diagnostic marker for ICP, but their concentrations are affected by environmental, dietary, and genetic factors, resulting in significant interindividual variation. This study aimed to develop and validate a clinically applicable nomogram for the early prediction of ICP risk. Methods: This prospective cohort study enrolled pregnant women attending prenatal care at Shanghai Public Health Clinical Center from June 2022 to June 2024. A total of 391 participants (184 ICP cases and 207 controls) were included. Clinical characteristics and serum biomarker concentrations were collected. Univariate analysis, LASSO regression, and multivariate logistic regression were used to identify independent predictors for ICP risk, which were then incorporated into a nomogram. Multicollinearity was assessed using the variance inflation factor (VIF), with VIF < 2 indicating its absence. Internal validation was performed using bootstrap resampling with 1,000 replicates. Model performance was evaluated for discrimination (AUC), calibration (Hosmer–Lemeshow test with calibration curve), and clinical utility (decision curve analysis and clinical impact curve). Results: Seven independent predictors for ICP risk were identified via multivariate logistic regression (all P < 0.05): age ≥ 35 years (OR = 5.276, 95% CI 1.155–24.727), ALT (OR = 1.082, 95% CI 1.023–1.163), TG (OR = 4.538, 95% CI 2.236–10.723), Apo A (OR = 0.001, 95% CI 0.000–0.089), FIB (OR = 3.460, 95% CI 1.288–10.726), sICAM-1 (OR = 1.067, 95% CI 1.036–1.112), and PLTP (OR = 1.001, 95% CI 1.000–1.001). Apo A was negatively associated with ICP, while the other six factors showed positive associations. No multicollinearity was detected (all VIF < 2). The nomogram exhibited excellent discrimination (AUC = 0.991) and optimal calibration (Hosmer-Lemeshow test: χ² = 0.991, P = 0.998). Decision curve analysis confirmed its clinical utility. Conclusion: The nomogram provides a reliable tool for the individualized risk assessment of ICP. It enables early identification of high-risk pregnant women, facilitates the optimization of perinatal monitoring and intervention strategies and may thereby improve maternal and neonatal outcomes.

Background: The preS1 region of the HBV large surface protein binds sodium taurocholate cotransporting polypeptide (NTCP) to mediate viral entry, but the serological and functional relevance of circulating preS1 antigen and anti-preS1 antibodies in chronic hepatitis B (CHB) is not well defined. Objective: To characterise serum preS1 antigen and anti-preS1 IgG across CHB and evaluate their virological and functional significance. Design: ELISAs for preS1 antigen and anti-preS1 IgG were developed and applied to 549 individuals with chronic HBV infection, 107 individuals with functional cureand 110 vaccinated healthy controls. Neutralising activity was assessed using HBV and hepatitis D virus (HDV) infection models, preS1-NTCP competitive assays and peptide microarrays. Baseline anti-preS1 IgG was evaluated in 51 patients discontinuing nucleos(t)ide analogue (NUC) therapy. Results: PreS1 antigen was highest in hepatitis B e antigen (HBeAg)-positive patients and correlated with HBV DNA, hepatitis B surface antigen and HBeAg. Anti-preS1 IgG was detectable across disease phases, inversely associated with HBV DNA, positively correlated with alanine aminotransferase/aspartate aminotransferase and remained detectable in some individuals with functional cure. Sera with low preS1 and high anti-preS1 IgG showed potent HBV/HDV neutralising activity by inhibiting viral entry and blocking preS1-NTCP binding. Dominant linear epitopes mapped to preS1 regions involved in NTCP binding. High baseline anti-preS1 IgG was associated with sustained virological suppression after NUC withdrawal. Conclusion: PreS1 antigenaemia and anti-preS1 IgG exhibit distinct serological patterns linked to HBV replication and endogenous viral control. High anti-preS1 IgG confers potential neutralising activity and associates with sustained off-therapy suppression, highlighting its functional relevance as an indicator of host viral control.

Cytomegalovirus (CMV) pneumonia presents diagnostic challenges in AIDS patients, as plasma monitoring often fails to reflect pulmonary viral burden. This retrospective study evaluated the prognostic value of bronchoalveolar lavage fluid (BALF) CMV DNA loads in 189 AIDS patients with pulmonary infections and CD4⁺ T cell counts < 200 cells/μL. CMV DNA in BALF and plasma was quantified to analyze associations with immune status and 90-day all-cause mortality. CMV detection was significantly more frequent in BALF (49.7%) than plasma (26.6%), indicating viral compartmentalization. An optimal BALF cutoff of 10,000 copies/mL was established for mortality prediction. Patients exceeding this threshold exhibited significantly lower CD4⁺ counts, increased mechanical ventilation requirements (34.4% vs. 11.5%), and prolonged hospital stays. Crucially, a BALF CMV load > 10,000 copies/mL was identified as an independent predictor of 90-day mortality (adjusted odds ratio = 3.78; 95% CI: 1.12–12.71). In conclusion, pulmonary CMV replication is prevalent and often compartmentalized in AIDS patients. A BALF CMV DNA load exceeding 10,000 copies/mL serves as a biomarker of profound immunosuppression and independently predicts poor clinical outcomes, highlighting the necessity of quantitative BALF monitoring for risk stratification.

The mechanism underlying the differential cytotoxicity of curcumin in various cancer types, however, remains largely unclear. The aims of this study is to examine the concentration- and time-related effects of curcumin on two different breast cancer cells, MCF-7 and MDA-MB-231, and investigated the functional changes induced by curcumin treatment, as well as their relationship to the PI3K/Akt-SKP2-Cip/Kips pathway. First, WST-1 and clonogenic assay were performed to determine the cytotoxicity of curcumin in MCF-7 and MDA-MB-231 cells. Then, the expression of CDK interacting protein/Kinase inhibitory protein (Cip/Kips) members (p27, p21 and p57) and S-phase kinase-associated protein-2 (SKP2) was investigated by QRT PCR and Western Blotting. Curcumin’s effect on PI3K (phosphatidylinositol 3-kinase) /Akt and its substrates Foxo1 and Foxo3a were then studied by Western Blotting. Small interfering RNAs (siRNAs) targeting SKP2 was used to explore the relationship between SKP2 and Cip/Kips members. Finally, WST-1 assay was tested to explore the concomitant treatment with curcumin and the inhibition of PKB or SKP2 signaling on curcumin sensitivity in MCF-7 and MDA-MB-231 cells. We demonstrated MCF-7 and MDA-MB-231 cells exhibited differential responses to curcumin by WST-1 and clonogenic assay (MDA-MB-231 cells was sensitive, and MCF-7 cells was resistant), which were found to be related to the differential curcumin-mediated regulation of SKP2-Cip/Kips (p21 and p27 but not p57) signaling. The differential cellular responses were further linked to the converse effects of curcumin on PI3K/Akt and its substrates Foxo1 and Foxo3a. Importantly, PI3K inhibitor wortmannin could counteract both curcumin-induced phosphorylation of Akt and up-regulation of SKP2 in MCF-7 cells. Subsequent WST-1 assay demonstrated concomitant treatment with curcumin and wortmannin or SKP2 siRNA not only further augmented curcumin sensitivity in MDA-MB-231 cells but also overcame curcumin resistance in MCF-7 cells. Our study established PI3K/Akt-SKP2-Cip/Kips signaling pathway is involved in the mechanism of action of curcumin and revealed that the discrepant modulation of this pathway by curcumin is responsible for the differential susceptibilities of these two cell types to curcumin.