This study investigates the neuroprotective potential of extracellular vesicles (EVs) delivering quercetin-3-O-β-d-glucuronic acid (QG-EVs) in cerebral ischemia-reperfusion injury (CIRI). Targeted brain delivery of QG-EVs was confirmed, with neuron cells identified as pivotal in modulating CIRI through single-cell RNA sequencing (scRNA-seq). Activating transcription factor 4 (Atf4) was highlighted as a critical regulatory factor, and in vitro studies revealed that silencing Atf4 diminished the neuroprotective effects of QG-EVs, increasing oxidative stress levels and neuronal apoptosis. In a CIRI mouse model, the knockdown of Atf4 attenuated the protective outcomes provided by QG-EVs, further affirming the role of Atf4 in mediating neuroprotection. Behavioral assessments and protein analysis showed that QG-EVs significantly reduced neuronal damage and pro-apoptotic markers, while improving neurological function via Atf4 upregulation. The outcomes hint at the potential of QG-EVs as a beneficial therapeutic modality to mitigate neuronal damage in CIRI by enhancing Atf4 expression, highlighting its potential for improving ischemic stroke outcomes.

The density and composition of lymphocytes infiltrating colon tumors serve as predictive factors for the clinical outcome of colon cancer.Our previous studies highlighted the potent anti-cancer properties of the principal compounds found in Garcinia yunnanensis(YTE-17),attributing these effects to the regu-lation of multiple signaling pathways.However,knowledge regarding the mechanism and effect of YTE-17 in the prevention of colorectal cancer is limited.In this study,we conducted isobaric tags for relative and absolute quantification(iTRAQ)analysis on intestinal epithelial cells(IECs)exposed YTE-17,both in vitro and in vivo,revealing a significant inhibition of the Wnt family member 5a(Wnt5a)/c-Jun N-terminal kinase(JNK)signaling pathway.Subsequently,we elucidated the influence and mechanism of YTE-17 on the tumor microenvironment(TME),specifically focusing on macrophage-mediated T helper 17(Th17)cell induction in a colitis-associated cancer(CAC)model with Wnt5a deletion.Additionally,we performed the single-cell RNA sequencing(scRNA-seq)on the colonic tissue from the Wnt5a-deleted CAC model to characterize the composition,lineage,and functional status of immune mesenchymal cells during different stages of colorectal cancer(CRC)progression.Remarkably,our findings demon-strate a significant reduction in M2 macrophage polarization and Th17 cell phenotype upon treatment with YTE-17,leading to the restoration of regulatory T(Treg)/Th17 cell balance in azoxymethane(AOM)/dextran sodium sulfate(DSS)model.Furthermore,we also confirmed that YTE-17 effectively inhibited the glycolysis of Th17 cells in both direct and indirect co-culture systems with M2 macrophages.Notably,our study shed light on potential mechanisms linking the non-canonical Wnt5a/JNK signaling pathway and well-established canonical β-catenin oncogenic pathway in vivo.Specifically,we proposed that Wnt5a/JNK signaling activity in IECs promotes the development of cancer stem cells with β-catenin activity within the TME,involving macrophages and T cells.In summary,our study undergoes the po-tential of YTE-17 as a preventive strategy against CRC development by addressing the imbalance with the immune microenvironment,thereby mitigating the risk of malignancies.

Objective:To investigate the clinical and pathological characteristics of breast cancer with HER2 low expression.Methods:The data from 3 422 patients with invasive breast cancer which archived in Peking Union Medical College Hospital between April 2019 and July 2022 were retrospectively reviewed. Among them, 136 patients were treated with neoadjuvant chemotherapy. The tumor size, histological type, tumor differentiation, lymph node metastasis, Ki-67 index, the status of estrogen receptor, progesterone receptor and HER2 as well as pathological complete response (pCR) rate were collected.Results:The HER2 status of 3 286 patients without neoadjuvant therapy, 616 (616/3 286, 18.7%) score 0, 1 047 (1 047/3 286, 31.9%) score 1+, 1 099 (1 099/3 286,33.4%) score 2+ and 524 (524/3 286,15.9%) score 3+ by immunohistochemistry (IHC). Among the 1 070 IHC 2+ cases, 161 were classified as HER2 positive by reflex fluorescence in situ hybridization (FISH) assay. In our cohort, 1 956 cases of HER2-low (IHC 1+ and IHC 2+/FISH-) breast cancer were identified. Compared to the HER2 IHC 0 group, HER2-low tumors more frequently occurred in patients with hormone receptor (HR) positive ( P<0.001), Ki-67 index below 35% ( P<0.001), well or moderate differentiation ( P<0.001) and over the age of 50 ( P=0.008). However, there were no significant differences in histological type, tumor size, and lymph node metastasis between HER2-low and HER2 IHC 0 group. For patients who had neoadjuvant therapy, the pCR rate in the patients with HER2-low was lower than those with HER2 IHC 0 (13.3%, 23.9%), but there was no significant difference. Although HER2-low breast cancers showed a slightly lower pCR rate than HER2 IHC 0 tumors, no remarkable difference was observed between tumors with HER2-low and HER2 IHC 0 regardless of hormone receptor status. Conclusions:The clinicopathological features of HER2-low breast cancers are different from those with HER2 IHC 0. It is necessary to accurately distinguish HER2-low breast cancer from HER2 IHC 0 and to reveal whether HER2-low tumor is a distinct biological entity.

Objective To investigate the consistency between colonoscopic Boston bowel preparation score and the bowel cleanliness evaluated by pre-endoscopy naked eye faecal observation,so as to provide a guidance on bowel preparation.Methods From September 2018 to June 2019,convenience sampling method was used to select 150 inpatients who underwent colonoscopy in the Department of Gastroenterology of a tertiary hospital in Guangzhou as the research objects.Before colonoscopy,the compound polyethylene glycol electrolyte powder was taken orally according to the bowel preparation plan for cleaning the colorectum.Before the colonoscopic examination,the naked eye observation method by nurses was used to observe the transparency of the excreta to evaluate the cleanliness of colorectum.Then the colorectal cleanliness was evaluated by endoscope by the operator using the Boston bowel preparation assessment scale(BBPS)during colonoscopy.Results A total of 145 patients completed the study.The cleanliness of bowel preparation was 93.1%with the naked eye observation and 88.27%with colonoscopy.There was no significant difference between the two assessment methods in judging the effectiveness of bowel preparation(P<0.05).The sensitivity of naked eye observation in judging bowel preparation was 96.10%with a 29.41%of specificity.The positive predictive value was 91.11%,and the negative predictive value was 50%(Kappa=0.310,P<0.001).Conclusion The naked eye observation and evaluation method for bowel preparation has advantages in high sensitivity,low specificity,high positive predictive value and low negative predictive value.It can be used as a preliminary evaluation method for cleanliness of colorectum before colonoscopy.

Objective: To study the influence of procyanidins on the bonding strength of dentin bleached by carbamide peroxide to composite resin. Methods : By applying different treatments to dentin bonding interfaces, 120 human third molars were randomly divided into 12 groups (n = 10): W group (no bleaching+deionized water), Wa group (no bleaching+deionized water+aging), WT1 group (no bleaching+5% procyanidins for 1 min), WT1a group (no bleaching+5% procyanidins for 1 min+aging), WT2 group (no bleaching+5% procyanidins for 5 min), WT2a group (no bleaching+5% procyanidins for 5 min+aging), C group (carbamide peroxide+deionized water), Ca group (carbamide peroxide+deionized water+aging), CT1 group (carbamide peroxide+5% procyanidins for 1 min), CT1a group (carbamide peroxide+5% procyanidins for 1 min+aging), CT2 group (carbamide peroxide+5% procyanidins for 5 min), and CT2a group (carbamide peroxide+5% procyanidins for 5 min+aging). The bond strength to composite resin was measured by universal mechanical testing machine, microstructure and the nanoleakages were measured by scanning electron microscope immediately or after the thermal cycling aging test. Results: The immediate bond strength of the bleached groups pretreated with procyanidins for 1 min (P<0.001) and 5 min (P<0.001) was higher than that of Group C, and the difference was statistically significant. Meanwhile, there was no statistically significant difference between Group CT1 and Group CT2 (P = 1.000). After the thermal cycles, the bond strength of each group declined. The differences between Group W and Group Wa (P<0.001) and Group C and Group Ca (P<0.001) were statistically significant, but no significant differences between Group CT1 and Group CT1a (P = 0.052) or Group CT2 and Group CT2a (P = 0.053) were found. The main effects of “aging” (P<0.001), “bleaching” (P<0.001) and “procyanidins” (P<0.001) and the second-order interaction effects of “bleaching * procyanidins” (P = 0.008), “bleaching * aging” (P = 0.024), and “aging * procyanidins” (P<0.001) were statistically significant. SEM observations showed that the hybrid layers in Groups C, CT1 and CT2 were not clear, and the hybrid layers in Groups Ca, CT1a and CT2a were partially destroyed and disintegrated. Under backscattering mode, it was observed that there were a large number of silver nitrate particles in the hybrid layer of Group Ca, and the residual silver ions in the hybrid layer of Groups CT1a and CT2a were decreased. Conclusion Pretreatment with 5% procyanidins for 1 min can improve the immediate bond strength of dentin bleached by carbamide peroxide to composite resin and maintain bonding durability.

Enzalutamide (ENZ) is a second-generation androgen receptor (AR) antagonist used for the treatment of castration-resistant prostate cancer (CRPC) and reportedly prolongs survival time within a year of starting therapy. However, CRPC patients can develop ENZ resistance (ENZR), mainly driven by abnormal reactivation of AR signaling, involving increased expression of the full-length AR (ARfl) or dominantly active androgen receptor splice variant 7 (ARv7) and ARfl/ARv7 heterodimers. There is currently no efficient treatment for ENZR in CRPC. Herein, a small molecule LLU-206 was rationally designed based on the ENZ structure and exhibited potent inhibition of both ARfl and constitutively active ARv7 to inhibit PCa proliferation and suppress ENZR in CRPC. Mechanically, LLU-206 promoted ARfl/ARv7 protein degradation and decreased ARfl/ARv7 heterodimers through mouse double minute 2-mediated ubiquitination. Finally, LLU-206 exhibited favorable pharmacokinetic properties with poor permeability across the blood-brain barrier, leading to a lower prevalence of adverse effects, including seizure and neurotoxicity, than ENZ-based therapies. In a nutshell, our findings demonstrated that LLU-206 could effectively inhibit ARfl/ARv7-driven CRPC by dual-targeting of ARfl/ARv7 heterodimers and protein degradation, providing new insights for the design of new-generation AR inhibitors to overcome ARfl/ARv7-driven CRPC.

Background Paraquat (PQ) is a widely used herbicide that exerts neurotoxicity. The effects of PQ on neural stem cells (NSCs) through microglia mediated neuroinflammation remain limitedly studied. Objective To investigate the effects of PQ on the proliferation and neurogenesis of NSCs through neuroinflammation mediated by microglia. Methods Microglial cell lines (BV2 cells) and primary NSCs were used. BV2 cells were exposed to 0, 1, 3.3, 10, 33, and 100 μmol·L⁻¹ of PQ for 6 h followed by viability assessment. The highest PQ concentration that had no effect on cell viability was selected as the final exposure concentration (33 μmol·L⁻¹). To exclude the direct effect of PQ on NSCs, after the BV2 cells were cultured in complete medium containing 33 μmol·L⁻¹ PQ for 6 h, the BV2 culture medium was replaced by NSCs complete medium without PQ for 24 h. The concentration of interleukin-1β (IL-1β) in supernatant was detected by enzyme-linked immune sorbent assay. Besides, in order to detect the effects of IL-1β on NSCs proliferation and neurogenesis, NSCs isolated from hippocampus of adult mice were cultured in the supernatant obtained above and divided into four groups: control supernatant + control antibody, control supernatant + IL-1β neutralizing antibody (10 ng·mL⁻¹), PQ supernatant + control antibody, PQ supernatant + IL-1β neutralizing antibody (10 ng·mL⁻¹). Proportion of Ki67-positive NSCs was detected by flow cytometry (FCS) and immunofluorescence after 24 h culture, and neurogenesis was detected by FCS and immunofluorescence after 3-7 d of culture. Results The IL-1β concentration in the supernatant of BV2 cells was significantly increased after the 33 μmol·L⁻¹ PQ exposure compared with the control group (t=3.020, P<0.05). After the NSCs were cultured with the supernatant of PQ-treated BV2 cells, the proportion of Ki67-positive NSCs (t=9.129, P<0.01) and the proportion of newborn neurons (t=4.638, P<0.01) were significantly decreased compared to the control group. After neutralizing IL-1β, the proportion of Ki67-positive NSCs (t=22.05, P<0.01) and the proportion of newborn neurons (t=11.09, P<0.01) were significantly higher than those in the un-neutralized group. The results of immunofluorescence detection also showed that after neutralizing IL-1β secreted by 33 μmol·L⁻¹ PQ-treated BV2 cells, the number of Ki67-positive NSCs and the number of newborn neurons were significantly higher than those in the un-neutralized group. Conclusion The secretion of IL-1β by microglia is increased after PQ treatment, resulting in a decrease in the proliferation and neurogenesis of NSCs. These results suggest that neuroinflammation is involved in NSCs damage caused by PQ.

Objective: To investigate the effects of XIAOJI Decoction combined with FOLFOX chemotherapy on serum cytokine expression profile in patients with advanced colorectal carcinoma by liquid chip technology. Methods: Fourteen patients with advanced colorectal carcinoma, who met the inclusion criteria and were treated in the Department of Oncology, Higher Education Mega Center Hospital of Guangdong Provincial Hospital of Traditional Chinese Medicine during January 1, 2018 and December 31, 2018 were retrospectively analyzed in this study. The patients were divided into chemotherapy group (n=7, treated with 5-Fluorouracil + Calcium Folic Acid+Oxaliplatin (FOLFOX)) and combined treatment group (n=7, treated with XIAOJI Decoction + FOLFOX) according to therapeutic scheme. The curative efficacy was evaluated after 6 treatment courses. The expression profile of cytokines in blood serum of patients was examined by liquid chip technology after every 2 courses. Results: Fourteen patients received a total of 84 cycles of therapy. Survival analyses showed that the progress-free survival time (PFS) and overall survival time (OS) of two groups couldn't be compared due to insufficient samples, although the combined treatment group had longer PFS (10 months vs 6 months) and OS (17 months vs 12 months) than the chemotherapy group.As to adverse reactions, the rates of leucopenia, diarrhea, nausea, peripheral neuritis and alopecia in two groups were comparable, while the severity in combined treatment group were lighter than that in chemotherapy group. In comparison with the combined treatment group, concentrations of serum BDNF and IL-2 were statistically higher in the chemotherapy group (P<0.05). By comparing the cytokine concentrations at different collection time points before and after the treatment, it showed that the concentration of serum IL-2 in chemotherapy group was higher than that in combined treatment group after 2 courses of treatment (P<0.05). In total, there were 19 cytokines showed a tendency to be higher in combined treatment group than chemotherapy group during different treatment periods. Conclusion: Combined treatment of XIAOJI Decoction with FOLFOX for advanced colorectal carcinoma is a treatment option worth exploring, and liquid chip analysis showed that the mechanism may be related to the reduction of serum LI-2 and BDNF levels in patients.

Objective The aim of this study was to investigate the mechanism of sanguinarine(SANG)on the inhibitory pro-liferation in ovarian cancer SKOV3 cells. Methods CCK-8 assay was used to detect proliferation of SKOV3 cells. Flow cytometry was used to detect the effect of SANG on apoptosis in SKOV3 cells. The spectrophotometer was used to detect the production of reac-tive oxygen species(ROS)by SANG. The mouse ovarian cancer xenograft model was used to detect the inhibitory effect of SANG on tumor growth. Results SANG promoted apoptosis in SKOV3 cells in a dose-and time-dependent manner. The SANG-induced ap-optosis was associated with the production of ROS,Activated the c-Jun-N-terminal kinase( JNK) and nuclear factor-κB( NF-κB)signaling pathways. In mouse model of ovarian cancer xenografts,after intravenous injection of mice with SANG,SANG was signifi-cantly inhibited the growth of ovarian cancer xenografts when compared to the control group. SANG also significantly induced apoptosis in ovarian cancer xenografts. Conclusion SANG can significantly inhibit the proliferation of ovarian cancer SKOV3 cells,induce ap-optosis,increase the production of ROS,and inhibit the growth of ovarian cancer.

Artificial joint replacement is an important means for the treatment of severe joint end-stage diseases such as hip and knee joint,which has obtained satisfactory clinical efficacy,but the postoperative periprosthetic osteolysis (PPO),which is mediated by wear particles,restricts the long-term effect of artificial joints.It is found that wear particles increase the expression of cytokines and inflammatory factors by stimulating the cells around the prosthesis,activate different signaling pathways,promote the imbalance between bone formation mediated by local osteoblasts and bone resorption mediated by osteoclast so as to lose of the local bone mass,and eventually produce osteolysis and aseptic loosening.This article reviews the different signal pathways activated by wear particles in recent years,in order to explore the pathogenesis of PPO and to open up new avenues for its prevention and treatment.