ObjectiveType I interferon (IFN-I) signaling is essential for antiviral innate immunity, yet its sustained or excessive activation contributes to the pathogenesis of several autoimmune diseases and interferonopathies, such as systemic lupus erythematosus and Aicardi-Goutières syndrome. Current strategies targeting this pathway, exemplified by JAK inhibitors, act mainly on downstream signal transduction and provide limited direct control over upstream IFN-I production, while also carrying the risk of broad immunosuppression. Phyllanthus emblica L. has long been used in traditional medicine for inflammatory disorders, but the bioactive constituent responsible for its regulation of IFN-I signaling and the underlying molecular mechanism have not been clearly defined. This study aimed to identify the active anti-inflammatory component of P. emblica and to characterize its mechanism of action on the IFN-I pathway in macrophages. MethodsActive components ofP. emblica and their candidate targets were screened by network pharmacology using the TCMSP and DrugBank databases (oral bioavailability≥30%, drug-likeness≥0.18) and intersected with inflammation-related genes retrieved from public databases. The predicted interaction between EGCG and IFN-I pathway proteins (TBK1, IRF3, STAT1) was evaluated by molecular docking, with BX795 and GSK8612 used as reference TBK1 inhibitors. Mechanistic experiments were performed in THP-1-derived macrophages and primary bone marrow-derived macrophages (BMDM). Upstream signaling was activated by transfection of the nucleic acid analogs poly(I∶C) and poly(dA∶dT) or by lipopolysaccharide (LPS) stimulation, whereas downstream signaling was activated by exogenous IFN-β. An siRNA-mediated TREX1 knockdown model was used to mimic endogenous nucleic acid-driven interferonopathy. Expression of IFN-β1 and interferon-stimulated genes (ISGs) was measured by RT-qPCR, protein phosphorylation by Western blot, and IFN-β secretion by ELISA. Cellular thermal shift assay (CETSA) and drug affinity responsive target stability (DARTS) were used to probe the interactionbetween EGCG and IRF3. ResultsNetwork pharmacology identified (-)-epigallocatechin-3-gallate (EGCG) as a candidate IFN-I-suppressive constituent of P. emblica, with predicted binding to TBK1, IRF3, and STAT1. Molecular docking yielded binding energies of -9.2, -7.2, and -8.2 kcal/mol for TBK1, IRF3, and STAT1, respectively, indicating an affinity for TBK1 comparable to that of the reference inhibitors BX795 (-5.7 kcal/mol) and GSK8612 (-6.4 kcal/mol). EGCG suppressed IFN-β1 and ISG mRNA expression under poly (I∶C), poly (dA∶dT), and LPS stimulation in both THP-1 macrophages and BMDM. At the protein level, EGCG reduced the phosphorylation of TBK1 and IRF3 without affecting the levels of the upstream sensors cGAS and RIG-I, and lowered IFN-β secretion in a concentration-dependent manner. CETSA and DARTS showed that EGCG did not enhance the thermal stability or protease resistance of IRF3, indicating that its effect on IRF3 is indirect. Following IFN-β stimulation, prolonged EGCG treatment reduced STAT1 phosphorylation in a time-dependent manner without an apparent change in IRF9, and partially attenuated ISG transcription; this effect was not monotonicly concentration-dependent, and CXCL10 showed the most consistent suppression. In TREX1-knockdown cells, the elevated mRNA levels of ISG15, ISG56, and CXCL10 were reduced by EGCG. ConclusionEGCG suppresses IFN-I responses by concurrently inhibiting TBK1-IRF3-dependent IFN‑β production and JAK-STAT1-mediated downstream transcription. These in vitro findings provide a mechanistic basis for the anti-inflammatory use of P. emblica in traditional medicine and identify EGCG as a candidate for further evaluation in interferon-driven autoimmune disease models.

OBJECTIVE: To investigate the association between ABO blood types and gestational diabetes mellitus (GDM) risk. METHODS: A prospective birth cohort study was conducted. ABO blood types were determined using the slide method. GDM diagnosis was based on a 75-g, 2-h oral glucose tolerance test (OGTT) according to the criteria of the International Association of Diabetes and Pregnancy Study Groups. Logistic regression was applied to calculate the odds ratios ( ORs) and 95% confidence intervals ( CIs) between ABO blood types and GDM risk. RESULTS: A total of 30,740 pregnant women with a mean age of 31.81 years were enrolled in this study. The ABO blood types distribution was: type O (30.99%), type A (26.58%), type B (32.20%), and type AB (10.23%). GDM was identified in 14.44% of participants. Using blood type O as a reference, GDM risk was not significantly higher for types A ( OR = 1.05) or B ( OR = 1.04). However, women with type AB had a 19% increased risk of GDM ( OR = 1.19, 95% CI = 1.05-1.34; P < 0.05), even after adjusting for various factors. This increased risk for type AB was consistent across subgroup and sensitivity analyses. CONCLUSION The ABO blood types may influence GDM risk, with type AB associated with a higher risk. Incorporating it-either as a single risk factor or in combination with other known factors-could help identify individuals at risk for GDM before or during early pregnancy.
Humans ; Female ; Pregnancy ; Diabetes, Gestational/etiology* ; ABO Blood-Group System ; Adult ; Prospective Studies ; Risk Factors ; Young Adult

AIM To investigate the repair effects of tauroursodeoxycholic acid(TUDCA)combined with bone marrow mesenchymal stem cells(BMSCs)transplantation on spinal cord injury(SCI)in rats.METHODS The rats were randomly divided into the sham operation group,the model group,the TUDCA group,the BMSCs transplantation group and the combination therapy of TUDCA and BMSCs transplantation group,with the SCI rat model established by Allen's method.The next day after modeling,the rats of TUDCA and combination therapy groups were given 200 mg/kg TUDCA by gavage.On the 3rd day after modeling,rats in BMSCs transplantation group and combination therapy group were injected with 1 mL tuned bone marrow BMSCs(the 3rd generation,1× 106/mL)via tail vein.Rats in the sham operation group and the model group were given gastric perfusion of normal saline and injection of 1 mL PBS through tail vein.On the 3rd,7th and 14th day after modeling,the rats had their motor function of hind limbs observed and BBB score determined.After the corresponding drug administration,the rats had their movement track of hind limbs recorded by footprint experiment;their the protein expressions of IL-6,IL-10,Arg-1,PI3K and Akt in spinal cord tissue detected by Western blot;their pathological changes of spinal cord tissue observed by HE staining and Nissl staining;and their expressions of MAP2,GAP43 and GFAP detected by immunofluorescence staining.RESULTS Compared with the model group,the groups intervened with TUDCA,or BMSCs transplantation,or combination therapy shared improved hind limb function and spinal cord histomorphology(P＜0.05);increased fluorescence intensity of MAP2 and GAP43,and protein expressions of IL-10,Arg-1,p-PI3K and p-Akt(P＜0.05);decreased fluorescence intensity of GFAP and IL-6 protein expressions(P＜0.05);among which the combination therapy group took the lead(P＜0.05).CONCLUSION The combination therapy of TUDCA and BMSCs transplantation may restore the function of the rat model of SCI by reducing inflammatory reaction,alleviating secondary injury,and promoting axon and myelin regeneration via PI3K/Akt signaling pathway.

Objective:This study constructed a mouse model carrying the human SNCA gene with E46K and A53T double mutations through transgenic technology,providing a suitable experimental animal model for drug screening,safety evaluation,pathogenesis research of Parkinson's disease,and studies on neurodegenerative diseases associated with abnormal α-synuclein aggregation.Methods:Using transgenic techniques,we introduced the human SNCA gene with the E46K and A53T mutations into the C57BL/6J mouse genome.These mutations are associated with familial PD and are known to promote α-Synuclein aggregation and neurotoxicity.The resulting B6-hSNCA E46K/A53T transgenic mice were systematically evaluated through behavioral tests to assess motor dysfunction,immunohistochemistry to characterize α-Synuclein pathology,and Western blotting to quantify molecular changes.Additionally,the therapeutic potential of glial cell line-derived neurotrophic factor(GDNF)delivered via adeno-associated virus(AAV)was assessed.Results:The B6-hSNCA E46KA53T double-mutant mice exhibitα-Synuclein aggregation in brain regions such as the cortex,brainstem,and cerebellum starting from 1-months-old.Phosphorylated α-Synuclein protein at serine 129 is detected in regions including the cortex and hippocampus.By 2-months-old,these mice begin to show significant declines in limb strength and motor coordination,as evidenced by grip strength and rotarod tests,displaying motor impairments reminiscent of Parkinson's disease.From 3-months-old,high-performance liquid chromatography(HPLC)reveals a reduction in dopamine and its metabolites in the striatum.Following treatment with AAV-GDNF injection,the mice demonstrate partial improvement in motor behaviors,as observed in rotarod and grip strength behavioral tests.Conclusion:The B6-hSNCA E46KA53T double-mutant mouse model effectively simulates the onset and progression of Parkinson's disease,demonstrating high clinical relevance.This model not only serves as a valuable tool for investigating the pathogenesis of Parkinson's disease but also provides a critical experimental platform for screening and safety evaluation of drugs targeting abnormal α-Synuclein aggregation.It holds significant potential for advancing the development of early diagnostic methods and targeted therapeutic strategies for Parkinson's disease.

Objective:To investigate the distribution of CGG repeat numbers in the FMR1 gene among reproductive-age women in China, providing data reference for carrier screening and genetic counseling of Fragile X syndrome. Methods:This cross-sectional study recruited 16 610 reproductive-age women from 12 medical institutions between July 2022 and October 2023. Peripheral venous blood samples (3 mL) were collected, and genomic DNA was extracted. The number of CGG repeats in the FMR1 gene was determined using the triplet-primed polymerase chain reaction (TP-PCR) combined with capillary electrophoresis technology. Statistical analyses were performed to assess the prevalence and distribution of CGG repeat expansions. Results:Among 16 610 women of childbearing age, 5 684 (34.220%) women had the same number of CGG repeats in the two alleles of FMR1 gene, and 10 926 (65.780%) women had different numbers of repeats in the two alleles. Among the 33 220 FMR1 alleles in 16 610 women of reproductive age, the most common CGG repeat numbers were 29 [48.645% (16 160/33 220)] and 30 [26.276% (8 729/33 220)], while the most frequent CGG genotype was CGG 29/29 [24.726% (4 107/16 610)]. The CGG repeat numbers of FMR1 gene were normal in 16 498 women (99.326%). Among the 112 women (0.674%) with CGG repeat abnormities, 96 (0.578%) women were classified as intermediate carriers, 15 (0.090%) as premutation carriers, and 1 (0.006%) as a full mutation carrier, whose CGG genotype was (36, >200). Conclusion:In the general reproductive-age female population in China, the normal CGG repeat numbers of the FMR1 gene account for 99.326%, while the intermediate carrier rate is 0.578%, and the combined carrier rate of the premutation and full mutation types is 0.096%.

OBJECTIVE To screen endogenous differential metabolites in mice that die from chlor-promazine(CPZ)poisoning and investigate the detoxification mechanism of triheptanoin(TriHep)against CPZ-induced lethality.METHODS Mice were randomly divided into the following groups(half male and half female):normal control,CPZ 2.5LD50,CPZ LD50 intoxication(CPZI),CPZ LD50 death(CPZD),TriHep-control,and TriHep-intervention(TriHep+CPZ LD50).The CPZ 2.5LD50,CPZI and CPZD groups were intragastrically given a corresponding dose of CPZ,respectively.The TriHep-control group and the TriHep-intervention group were intragastrically given saline and CPZ LD50 respectively before being intragastrically given TriHep(3 μL·g-1)10 min later.Plasma samples from the CPZ 2.5LD50 group and normal control group were analyzed using liquid chromatography-tandem mass spectrometry(LC-MS/MS)for metabolite identification and quantification.MetaboAnalyst 5.0 was employed to perform principal component analysis(PCA),orthogonal partial least squares-discriminant analysis(OPLS-DA),and metabolic pathway analysis to screen and identify differential metabolites.More comparisons were made of the levels of differential metabolites in plasma between the normal control,CPZI,CPZD,TriHep-intervention,and TriHep-control groups.RESULTS In the PCA score plot,metabolomic samples from the CPZ 2.5LD50 group and normal control group showed clear separation,indicating distinct clus-tering patterns.Primary screening under three conditions,including P＜0.05,variable importance in projec-tion(VIP)score≥ 1 and fold change(FC)≥1.5 or ≤0.67 for a comparison of CPZ 2.5LD50 group with normal control group 28 metabolites were identified.Following quantitative enrichment and structural identifica-tion,three significantly differential metabolites were confirmed:acetylcarnitine,propionylcarnitine,and succinic acid.Compared with the normal control group,both CPZI and CPZD groups showed signifi-cantly decreased plasma levels of acetylcarnitine and propionylcarnitine,while the succinic acid content was markedly increased in the CPZD group.In the TriHep control group,levels of acetylcarnitine and succinic acid were significantly elevated,with no significant change in propionylcarnitine levels.Com-pared with the CPZI group,the CPZD group showed a significant increase in plasma succinic acid levels,but no significant change was observed in the acetylcarnitine content.The TriHep-intervention group demonstrated metabolite profiles(all the three differential metabolites)similar to those in the CPZI group,with significantly reduced propionylcarnitine and succinic acid concentrations compared to the CPZD group.CONCLUSION In the early stage of CPZ intoxication,TriHep can alleviate CPZ poisoning via acetylcarnitine,which can stabilize the level of succinic acid in plasma via indirect succinic acid replenishment.

Objective:To investigate the distribution of CGG repeat numbers in the FMR1 gene among reproductive-age women in China, providing data reference for carrier screening and genetic counseling of Fragile X syndrome. Methods:This cross-sectional study recruited 16 610 reproductive-age women from 12 medical institutions between July 2022 and October 2023. Peripheral venous blood samples (3 mL) were collected, and genomic DNA was extracted. The number of CGG repeats in the FMR1 gene was determined using the triplet-primed polymerase chain reaction (TP-PCR) combined with capillary electrophoresis technology. Statistical analyses were performed to assess the prevalence and distribution of CGG repeat expansions. Results:Among 16 610 women of childbearing age, 5 684 (34.220%) women had the same number of CGG repeats in the two alleles of FMR1 gene, and 10 926 (65.780%) women had different numbers of repeats in the two alleles. Among the 33 220 FMR1 alleles in 16 610 women of reproductive age, the most common CGG repeat numbers were 29 [48.645% (16 160/33 220)] and 30 [26.276% (8 729/33 220)], while the most frequent CGG genotype was CGG 29/29 [24.726% (4 107/16 610)]. The CGG repeat numbers of FMR1 gene were normal in 16 498 women (99.326%). Among the 112 women (0.674%) with CGG repeat abnormities, 96 (0.578%) women were classified as intermediate carriers, 15 (0.090%) as premutation carriers, and 1 (0.006%) as a full mutation carrier, whose CGG genotype was (36, >200). Conclusion:In the general reproductive-age female population in China, the normal CGG repeat numbers of the FMR1 gene account for 99.326%, while the intermediate carrier rate is 0.578%, and the combined carrier rate of the premutation and full mutation types is 0.096%.

OBJECTIVE To screen endogenous differential metabolites in mice that die from chlor-promazine(CPZ)poisoning and investigate the detoxification mechanism of triheptanoin(TriHep)against CPZ-induced lethality.METHODS Mice were randomly divided into the following groups(half male and half female):normal control,CPZ 2.5LD50,CPZ LD50 intoxication(CPZI),CPZ LD50 death(CPZD),TriHep-control,and TriHep-intervention(TriHep+CPZ LD50).The CPZ 2.5LD50,CPZI and CPZD groups were intragastrically given a corresponding dose of CPZ,respectively.The TriHep-control group and the TriHep-intervention group were intragastrically given saline and CPZ LD50 respectively before being intragastrically given TriHep(3 μL·g-1)10 min later.Plasma samples from the CPZ 2.5LD50 group and normal control group were analyzed using liquid chromatography-tandem mass spectrometry(LC-MS/MS)for metabolite identification and quantification.MetaboAnalyst 5.0 was employed to perform principal component analysis(PCA),orthogonal partial least squares-discriminant analysis(OPLS-DA),and metabolic pathway analysis to screen and identify differential metabolites.More comparisons were made of the levels of differential metabolites in plasma between the normal control,CPZI,CPZD,TriHep-intervention,and TriHep-control groups.RESULTS In the PCA score plot,metabolomic samples from the CPZ 2.5LD50 group and normal control group showed clear separation,indicating distinct clus-tering patterns.Primary screening under three conditions,including P＜0.05,variable importance in projec-tion(VIP)score≥ 1 and fold change(FC)≥1.5 or ≤0.67 for a comparison of CPZ 2.5LD50 group with normal control group 28 metabolites were identified.Following quantitative enrichment and structural identifica-tion,three significantly differential metabolites were confirmed:acetylcarnitine,propionylcarnitine,and succinic acid.Compared with the normal control group,both CPZI and CPZD groups showed signifi-cantly decreased plasma levels of acetylcarnitine and propionylcarnitine,while the succinic acid content was markedly increased in the CPZD group.In the TriHep control group,levels of acetylcarnitine and succinic acid were significantly elevated,with no significant change in propionylcarnitine levels.Com-pared with the CPZI group,the CPZD group showed a significant increase in plasma succinic acid levels,but no significant change was observed in the acetylcarnitine content.The TriHep-intervention group demonstrated metabolite profiles(all the three differential metabolites)similar to those in the CPZI group,with significantly reduced propionylcarnitine and succinic acid concentrations compared to the CPZD group.CONCLUSION In the early stage of CPZ intoxication,TriHep can alleviate CPZ poisoning via acetylcarnitine,which can stabilize the level of succinic acid in plasma via indirect succinic acid replenishment.

Objective:This study constructed a mouse model carrying the human SNCA gene with E46K and A53T double mutations through transgenic technology,providing a suitable experimental animal model for drug screening,safety evaluation,pathogenesis research of Parkinson's disease,and studies on neurodegenerative diseases associated with abnormal α-synuclein aggregation.Methods:Using transgenic techniques,we introduced the human SNCA gene with the E46K and A53T mutations into the C57BL/6J mouse genome.These mutations are associated with familial PD and are known to promote α-Synuclein aggregation and neurotoxicity.The resulting B6-hSNCA E46K/A53T transgenic mice were systematically evaluated through behavioral tests to assess motor dysfunction,immunohistochemistry to characterize α-Synuclein pathology,and Western blotting to quantify molecular changes.Additionally,the therapeutic potential of glial cell line-derived neurotrophic factor(GDNF)delivered via adeno-associated virus(AAV)was assessed.Results:The B6-hSNCA E46KA53T double-mutant mice exhibitα-Synuclein aggregation in brain regions such as the cortex,brainstem,and cerebellum starting from 1-months-old.Phosphorylated α-Synuclein protein at serine 129 is detected in regions including the cortex and hippocampus.By 2-months-old,these mice begin to show significant declines in limb strength and motor coordination,as evidenced by grip strength and rotarod tests,displaying motor impairments reminiscent of Parkinson's disease.From 3-months-old,high-performance liquid chromatography(HPLC)reveals a reduction in dopamine and its metabolites in the striatum.Following treatment with AAV-GDNF injection,the mice demonstrate partial improvement in motor behaviors,as observed in rotarod and grip strength behavioral tests.Conclusion:The B6-hSNCA E46KA53T double-mutant mouse model effectively simulates the onset and progression of Parkinson's disease,demonstrating high clinical relevance.This model not only serves as a valuable tool for investigating the pathogenesis of Parkinson's disease but also provides a critical experimental platform for screening and safety evaluation of drugs targeting abnormal α-Synuclein aggregation.It holds significant potential for advancing the development of early diagnostic methods and targeted therapeutic strategies for Parkinson's disease.

AIM To investigate the repair effects of tauroursodeoxycholic acid(TUDCA)combined with bone marrow mesenchymal stem cells(BMSCs)transplantation on spinal cord injury(SCI)in rats.METHODS The rats were randomly divided into the sham operation group,the model group,the TUDCA group,the BMSCs transplantation group and the combination therapy of TUDCA and BMSCs transplantation group,with the SCI rat model established by Allen's method.The next day after modeling,the rats of TUDCA and combination therapy groups were given 200 mg/kg TUDCA by gavage.On the 3rd day after modeling,rats in BMSCs transplantation group and combination therapy group were injected with 1 mL tuned bone marrow BMSCs(the 3rd generation,1× 106/mL)via tail vein.Rats in the sham operation group and the model group were given gastric perfusion of normal saline and injection of 1 mL PBS through tail vein.On the 3rd,7th and 14th day after modeling,the rats had their motor function of hind limbs observed and BBB score determined.After the corresponding drug administration,the rats had their movement track of hind limbs recorded by footprint experiment;their the protein expressions of IL-6,IL-10,Arg-1,PI3K and Akt in spinal cord tissue detected by Western blot;their pathological changes of spinal cord tissue observed by HE staining and Nissl staining;and their expressions of MAP2,GAP43 and GFAP detected by immunofluorescence staining.RESULTS Compared with the model group,the groups intervened with TUDCA,or BMSCs transplantation,or combination therapy shared improved hind limb function and spinal cord histomorphology(P＜0.05);increased fluorescence intensity of MAP2 and GAP43,and protein expressions of IL-10,Arg-1,p-PI3K and p-Akt(P＜0.05);decreased fluorescence intensity of GFAP and IL-6 protein expressions(P＜0.05);among which the combination therapy group took the lead(P＜0.05).CONCLUSION The combination therapy of TUDCA and BMSCs transplantation may restore the function of the rat model of SCI by reducing inflammatory reaction,alleviating secondary injury,and promoting axon and myelin regeneration via PI3K/Akt signaling pathway.