Objective To investigate the immunomodulatory effects of Aedes albopictus salivary protein rAlb-34kDa-1 on macrophages (Raw264.7) and its impact on dengue virus type 2 (DENV-2) replication in Raw264.7 cells. Methods The prokaryotic expression plasmid pET32a(+)-rAlb-34kDa-1 was transfected into Escherichia coli (E.coli) BL21 (DE3), and salivary proteins were obtained according to the expression purification conditions established in the pre-laboratory stage and identified by SDS-PAGE and Western blot (WB). The toxic effects of rAlb-34kDa-1 protein in Raw264.7 cells were assessed by CCK8 assay. Raw264.7 cells were treated with rAlb-34kDa-1 at concentrations of 0.02, 0.2, and 2 μg/mL for 6, 12, and 24 h. The expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), chemokine ligand 2 (CCL2), interferon-β (IFN-β), and interferon-stimulated gene 15 (ISG15) within the cells was detected using real-time quantitative reverse transcription PCR (qRT-PCR). Enzyme-linked immunosorbent assay (ELISA) was used to measure IL-6 and TNF-α secretion in the cell culture supernatants. Immunofluorescence staining was used to observe the translocation of p65 into the nucleus of Raw264.7 cells following rAlb-34kDa-1 treatment, to analyze whether rAlb-34kDa-1 modulates the immune response in Raw264.7 cells through NF-κB signaling pathway. Cells were collected 6, 12, and 24 hours post-treatment under different experimental conditions (DENV-2 treatment group, salivary protein +DENV-2 co-incubation group) was measured using qRT-PCR, and the fluorescence intensity of J2 (dsRNA) in the cells was detected by flow cytometry (FCM) after 24 hours under different treatment factors. Results The results of SDS-PAGE and WB confirmed the successful acquisition of rAlb-34kDa-1 protein, and CCK8 analysis showed no cytotoxicity of this protein toward Raw264.7 cells. Both 0.2 μg/mL and 2 μg/mL rAlb-34kDa-1 induced the expression of IL-1β, IL-6, TNF-α, CCL2, IFN-β, and ISG15 in Raw264.7 cells. One hour after rAlb-34kDa-1 treatment, the fluorescence intensity of p65 in the nucleus of Raw264.7 cells was significantly enhanced. In Raw264.7 cells co-incubated with rAlb-34kDa-1 and DENV-2 for 12 hours, rAlb-34kDA-1 at a concentration of 2 μg/mL significantly increased the mRNA expression of DENV-2. After 24 hours of treatment, rAlb-34kDA-1 at concentrations of 0.02 μg/mL, 0.2 μg/mL, and 2 μg/mL all enhanced the mRNA expression of DENV-2 in Raw264.7 cells, and 2 μg/mL of rAlb-34kDa-1 co-incubated with DENV-2 significantly increased the J2 fluorescence signal in cells. Conclusion The rAlb-34kDa-1 protein can regulate the immune response in murine Raw264.7 cells and promote the replication of DENV-2 in these cells.

This study aims to delve into the influences and underlying mechanisms of Banxia Xiexin Decoction(BXD) on the proliferation, apoptosis, invasion, and migration of colon cancer cells. Firstly, the components of BXD in blood were identified by UPLC-MS/MS, and subsequently the content of these components were determined by HPLC. Then, different concentrations of BXD were used to treat both the normal intestinal epithelial cells(NCM460) and the colon cancer cells(HT29 and HCT116). The cell viability and apoptosis were examined by the cell counting kit-8(CCK-8) and flow cytometry, respectively. Western blot was employed to determine the expression of the apoptosis regulators B-cell lymphoma-2(Bcl-2) and Bcl-2-associated X(Bax). The cell wound healing assay and Transwell assay were employed to measure the cell migration and invasion, respectively. Additionally, Western blot was employed to determine the expression levels of epithelial-mesenchymal transition(EMT)-associated proteins, including epithelial cadherin(E-cadherin), neural cadherin(N-cadherin), and vimentin. The protein and mRNA levels of the factors in the poly(ADP-ribose) glycohydrolase(PARG)/poly(ADP-ribose) polymerase 1(PARP1)/nuclear factor kappa-B p65(NF-κB p65) signaling pathway were determined by Western blot and RT-qPCR, respectively. The results demonstrated that following BXD intervention, the proliferation of HT29 and HCT116 cells was significantly reduced. Furthermore, BXD promoted the apoptosis, enhanced the expression of Bcl-2, and suppressed the expression of Bax in colon cancer cells. At the same time, BXD suppressed the cell migration and invasion and augmented the expression of E-cadherin while diminishing the expression of N-cadherin and vimentin. In addition, BXD down-regulated the protein and mRNA levels of PARG, PARP1, and NF-κB p65. In conclusion, BXD may inhibit the malignant phenotypes of colon cancer cells by mediating the PARG/PARP1/NF-κB signaling pathway.
Colonic Neoplasms/pathology* ; Drugs, Chinese Herbal/pharmacology* ; Phenotype ; Signal Transduction/drug effects* ; Cell Proliferation/drug effects* ; Apoptosis ; Cell Movement/drug effects* ; Neoplasm Invasiveness ; HCT116 Cells ; Proto-Oncogene Proteins c-bcl-2/biosynthesis* ; Humans ; Poly (ADP-Ribose) Polymerase-1 ; Glycoside Hydrolases ; bcl-2-Associated X Protein ; NF-kappa B p50 Subunit

In hypoxic-ischemic brain damage (HIBD), the programmed cell death known as ferroptosis is significantly activated. Microglial cells demonstrate a high level of sensitivity to iron accumulation. Understanding how to regulate the dual role of microglia and transforming the microglial ferroptosis to a moderate and controllable process has considerable implications for the targeted treatment in HIBD. This paper serves as an overview of microglia-mediated ferroptosis in HIBD as a disease model. We discuss various aspects centered around microglia, including pathophysiological mechanisms, polarization and functions of microglia, molecular mechanisms of ferroptosis, signaling pathways, and therapeutic strategies. The review aims to provide a reference for studies of ferroptosis in microglia.
Microglia/physiology* ; Ferroptosis/physiology* ; Humans ; Animals ; Hypoxia-Ischemia, Brain/pathology* ; Signal Transduction

OBJECTIVES: To investigate the application value of thromboelastography (TEG) in assessing coagulation function in children with severe hemophilia A (HA) after emicizumab (EMI) therapy. METHODS: A retrospective analysis was performed on the activated partial thromboplastin time (APTT) and TEG testing results of 17 children with severe HA before and after EMI treatment at Shenzhen Children's Hospital from January 2023 to July 2024. Correlation analysis was conducted between coagulation factor VIII (FVIII) equivalent activity and reaction time (R value) measured by TEG. RESULTS: After EMI treatment, the mean bleeding rate for children with severe HA was 1.6 events per year, with 15 children (88%) without spontaneous bleeding or joint bleeding. The children with severe HA showed a significant reduction in APTT after EMI treatment (P<0.05), with a significantly shorter APTT than the normal control group (P<0.05). There was no correlation between APTT and FVIII equivalent activity after treatment (P>0.05). After EMI treatment, TEG parameters, including R value, kinetic time, alpha angle (α), maximum amplitude, clot strength, and coagulation index, shifted from a hypocoagulable state before treatment to a nearly normal state after treatment (P<0.05). The R value demonstrated a strong negative correlation with FVIII equivalent activity (r=-0.758, P<0.05). CONCLUSIONS The bleeding condition of children with severe HA can be effectively controlled after EMI treatment. Routine APTT testing cannot reflect true coagulation function, whereas TEG testing is clinically valuable in assessing the coagulation function of children with severe HA undergoing EMI treatment.
Humans ; Thrombelastography ; Hemophilia A/physiopathology* ; Male ; Child ; Antibodies, Bispecific/therapeutic use* ; Antibodies, Monoclonal, Humanized/therapeutic use* ; Blood Coagulation/drug effects* ; Child, Preschool ; Retrospective Studies ; Female ; Partial Thromboplastin Time ; Adolescent ; Infant

OBJECTIVES: Most dermatological conditions fall under visible skin diseases (VSDs), where lesions are exposed and readily seen, increasing patients' risk of experiencing external stigma from the public and specific professional groups (e.g., service providers). This stigma imposes psychological and social burdens that far exceed the psychological symptoms of the disease. To date, no systematic research has been conducted in China specifically on the external stigma associated with VSDs. Taking psoriasis, vitiligo, and acne as representative conditions, this study aims to explore the external stigma experienced by VSD patients across various social settings and to provide a scientific foundation for the development of measurement tools, quantitative research, and targeted interventions. METHODS: A purposive sample of 23 outpatients diagnosed with psoriasis, acne, or vitiligo was recruited from the Xiangya Hospital Dermatology Clinic of Central South University between December 2023 and July 2024. In-depth qualitative interviews were conducted. Data were analyzed using Mayring's qualitative content analysis and thematic analysis. Reporting followed the Consolidated Criteria for Reporting Qualitative Research guidelines. The interviews focused on the experience of external stigma across different social settings. RESULTS: Patients with VSDs reported experiencing external stigma in various contexts including family, community, recreational service venues, healthcare institutions, and others. The main motivation behind stigmatizing behaviors was disease avoidance (e.g., fear of contagion, aversion, social distancing). Stigmatization in school settings was also reported by patients with all 3 types of VSDs. Psoriasis patients reported stigma across all examined scenarios, while vitiligo and acne patients reported stigma in only some contexts. CONCLUSIONS Patients with VSDs experience significant external stigma, with psoriasis patients facing a higher burden compared to those with vitiligo or acne. The predominant stigma-driving factor is the public's desire to avoid disease, which underscores the need for public education to correct misconceptions about VSDs. External stigma from family, school, social networks, healthcare providers, and structural stigma should be the focus of policy and intervention efforts aimed at protecting the rights and well-being of patients with VSDs.
Humans ; Social Stigma ; Female ; Male ; Qualitative Research ; Acne Vulgaris/psychology* ; Skin Diseases/psychology* ; Adult ; Psoriasis/psychology* ; Vitiligo/psychology* ; Middle Aged ; China ; Young Adult

OBJECTIVES: To evaluate the therapeutic effect of gastrodin against hypoxic-ischemic brain damage (HIBD) in neonatal mice and explore the role of GPX4/SLC7A11/FTH1 signaling in mediating its effect. METHODS: Twenty-four 9- to 11-day-old C57BL/6J mice were randomized equally into 4 groups for sham operation, HIBD modeling by right common carotid artery ligation and subsequent exposure to hypoxia for 1 h, or gastrodin treatment at 100 or 200 mg/kg before and at 1 and 2 days after modeling. The mice then underwent neurological assessment (Zea-Longa scores), and the cerebral cortical penumbra tissue were collected for HE and Nissl staining, detection of ferroptosis biomarkers and protein expressions of GPX4, SLC7A11, and FTH1 with Western blotting and immunofluorescence co-localization, and observation of mitochondrial ultrastructure with electron microscopy. In cultured HT22 neuronal cells with oxygen-glucose deprivation (OGD) for 2 h, the effects of pretreatments with 0.5 mmol/L gastrodin, 10 μmol/L RSL3 (a GPX4 inhibitor), alone or in combination, were analyzed on expressions of ferroptosis-related proteins, cellular Fe²⁺, ROS, lipid peroxidation, MDA, and GSH levels, mitochondrial membrane potential (JC-1), and cell viability. RESULTS: Gastrodin treatment at the two doses both significantly ameliorated HIBD and neurological deficits of the mice, reduced mitochondrial damage and Fe²⁺, MDA and ROS levels, increased GSH level, and upregulated GPX4, SLC7A11, and FTH1 protein expressions. In HT22 cells, gastrodin pretreatment obviously attenuated OGD-induced ferroptosis and improved cell viability and mitochondrial function. Co-treatment with RSL3 potently abrogated the inhibitory effects of gastrodin on Fe²⁺, ROS, BODIPY-C11, and MDA levels and attenuated its protective effects on GSH level, cell viability, and mitochondrial membrane potential. CONCLUSIONS Gastrodin provides neuroprotective effects in neonatal mice with HIBD by suppressing neuronal ferroptosis via upregulating the GPX4/SLC7A11/FTH1 signaling pathway.
Animals ; Ferroptosis/drug effects* ; Hypoxia-Ischemia, Brain/drug therapy* ; Mice ; Mice, Inbred C57BL ; Signal Transduction/drug effects* ; Phospholipid Hydroperoxide Glutathione Peroxidase ; Glucosides/pharmacology* ; Animals, Newborn ; Benzyl Alcohols/pharmacology* ; Amino Acid Transport System y+/metabolism*

Objective To explore whether renal denervation (RDN) could improve the left ventricular ejection fraction (LVEF) of patients with heart failure (HF) on the basis of guideline-directed management and therapy (GDMT). Methods From January 1, 2023 to August 31, 2024, HF patients diagnosed as dilated cardiomyopathy (DCM) who underwent RDN in the Second Affiliated Hospital of Chongqing Medical University were retrospectively enrolled, all patients had received GDMT for at least three months but the LVEF remained below 55%. Parameters of transthoracic echocardiography (TTE) at baseline, during GDMT, and after RDN were compared to analyze whether RDN can further improve the LVEF of patients on the basis of GDMT. Results A total of 7 HF patients diagnosed with DCM were enrolled, the mean age was (52.86±9.86) years old, and 5(71.4%) were male. After an average of (9.29±8.06) months of GDMT, LVEF significantly increased from baseline (34.86%±10.22%) to (44.57%±5.59%, P=0.024).Three months after RDN, LVEF was further significantly improved (54.43%±9.05%, P=0.026). The average follow-up after RDN was (11.00±4.12) months. The LVEF remained stable (54.86%±7.10%, P=0.805), and no adverse events occurred in the patients. Conclusions RDN can further enhance the LVEF of HF patients on the basis of GDMT.

Objective To explore the relationship between the polymorphism of excision repair cross-complementation group 1 (ERCC1) C8092A locus and the efficacy and prognosis of platinum-based chemotherapy for lung cancer (LC), and to provide a theoretical basis for precision treatment of LC. Methods From January 2014 to October 2017, 120 patients from two tertiary hospitals in Haikou City, and with pathologically confirmed lung cancer treated with platinum-based chemotherapy were selected as the research objects. After informed consent was obtained, peripheral blood samples were collected for DNA extraction, and the genotype of ERCC1 C8092A locus was detected by mass spectrometry. WHO's Response Evaluation Criteria in Solid Tumours (RECIST) was used to judge patients' chemotherapy efficacy and patients' survival status was obtained by telephone follow-up and other means. Results Among the 120 LC patients, the genotype frequencies of ERCC1 C8092A locus were 67 cases of CC wildtype (55.8%), 45 cases of CA heterozygous type (37.5%), and 8 cases of AA rare mutation type (6.7%), which conformed to Hardy-Weinberg equilibrium (χ2=0.140, P>0.05). The total effective rate of chemotherapy was 32.5%, with the highest effective rate in patients with the CA genotype (42.2%) at the ERCC1 C8092A locus and the lowest in patients with the CC genotype (25.4%). The overall one-year survival rate was 68.3% and the three-year survival rate was 35.8%. The patients with ERCC1 C8092A AA genotype had the lowest survival rate, with a one-year survival rate of 50.0% and three-year survival rate of only 25.0%. However, there were no statistical differences in the overall survival rate among the three genotypes of carriers of ERCC1 C8092A (χ2=0.328, P=0.849). Conclusions The polymorphism of ERCC1 C8092A locus is associated with the efficacy of platinum-based chemotherapy for LC, and patients with CA genotype have the highest efficacy. The one-year and three-year survival rates of patients with CC genotype are significantly higher than those of CA and AA genotypes.

Objective To observe the efficacy and safety of Morinda officinalis oligosaccharides in the continuation treatment of mild and moderate depression.Methods An open,single-arm,multi-center design was adopted in our study.Adult patients with mild and moderate depression who had received acute treatment of Morinda officinalis oligosaccharides were enrolled and continue to receive Morinda officinalis oligosaccharides capsules for 24 weeks,the dose remained unchanged during continuation treatment.The remission rate,recurrence rate,recurrence time,and the change from baseline to endpoint of Hamilton Depression Scale(HAMD),Hamilton Anxiety Scale(HAMA),Clinical Global Impression-Severity(CGI-S)and Arizona Sexual Experience Scale(ASEX)were evaluated.The incidence of treatment-related adverse events was reported.Results The scores of HAMD-17 at baseline and after treatment were 6.60±1.87 and 5.85±4.18,scores of HAMA were 6.36±3.02 and 4.93±3.09,scores of CGI-S were 1.49±0.56 and 1.29±0.81,scores of ASEX were 15.92±4.72 and 15.57±5.26,with significant difference(P＜0.05).After continuation treatment,the remission rate was 54.59％(202 cases/370 cases),and the recurrence rate was 6.49％(24 cases/370 cases),the recurrence time was(64.67±42.47)days.The incidence of treatment-related adverse events was 15.35％(64 cases/417 cases).Conclusion Morinda officinalis oligosaccharides capsules can be effectively used for the continuation treatment of mild and moderate depression,and are well tolerated and safe.