ObjectiveTo investigate the migration and distribution characteristics of chemical constituents in blood and hippocampal tissues before and after vinegar processing of Citri Reticulatae Pericarpium Viride（CRPV）， and to explore the potential material basis and mechanisms underlying their regulatory effects on emotional disorders by comparing the effects of raw and vinegar-processed products of CRPV. MethodsUltra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS） was employed to characterize and identify the chemical constituents of raw and vinegar-processed products of CRPV extracts， as well as their migrating components in blood and hippocampal tissues after oral administration. Reference standards， databases， and relevant literature were utilized for compound annotation， with data processing performed using PeakView 1.2 software. Seventy male C57BL/6 mice were randomly divided into seven groups， including the blank group， model group， diazepam group（2.5 mg·kg^-1）， raw CRPV low/high dose groups（0.6， 1.2 g·kg^-1）， and vinegar-processed CRPV low/high dose groups（0.6， 1.2 g·kg^-1）， with 10 mice per group. Except for the blank group， all other groups underwent chronic restraint stress（2 h·d^-1） for 20 d. Each drug-treated group received oral administration at the predetermined dose starting 10 d after modeling， with a total treatment duration of 10 d. Following model-based drug administration， mice underwent open-field， forced swimming， and elevated plus maze tests. After anesthesia with isoflurane， whole brains were collected from each group of mice， and hippocampi were dissected. Reactive oxygen species（ROS） level in hippocampal tissues was quantified by enzyme-linked immunosorbent assay（ELISA）. Hematoxylin-eosin（HE） staining was used to observe hippocampal tissue morphology. Immunofluorescence was performed to detect neuronal nuclei（NeuN） and peroxisome proliferator-activated receptor alpha（PPARα） expressions in hippocampal tissue. Then， pharmacodynamic evaluations were conducted to assess the effects of raw and vinegar-processed CRPV on mood disorders， exploring the potential mechanisms. ResultsVinegar processing caused significant changes in the chemical composition of CRPV， with 18 components showing increased relative content and 35 components showing decreased relative content. The primary changes occurred in flavonoid compounds， including 20 flavonoids， 20 flavonoid glycosides， 3 triterpenes， 3 phenolic acids， 1 alkaloid， and 6 other compounds. Twenty-one components were detected in blood（15 methoxyflavones， 4 flavonoid glycosides， and 2 phenolic acids）， with 17 shared between raw and vinegar-processed CRPV. Seven components reached hippocampal tissues（all common to both forms）. In regulating emotional disorders， Vinegar-processed CRPV exhibited superior antidepressant-like effects compared to raw products. HE staining revealed that both treatments improved hippocampal neuronal morphology， particularly in the damaged CA1 and CA3 regions. Immunofluorescence and ELISA analyses demonstrated that both raw and vinegar-processed CRPV significantly modulated NeuN and PPARα expressions in hippocampal tissue while alleviating oxidative stress induced by excessive ROS（P<0.05）. ConclusionThe chemical composition of CRPV undergoes changes after vinegar processing， but the migrating components in blood and hippocampus are primarily methoxyflavonoids. These components may serve as the potential material basis for activating the PPARα pathway， thereby negatively regulating ROS generation in the hippocampus， reducing oxidative stress， and promoting the development of NeuN-positive neurons. These findings provide experimental evidence for enhancing quality standards， pharmacodynamic material research， and active drug development of raw and vinegar-processed CRPV.

Objective To elucidate the molecular mechanism of shugan formula in regulating intestinal flora in liver cancer using network pharmacology.Methods The active components and targets of shugan formula were screened using the TCMSP and BATMAN-TCM databases.Targets associated with liver cancer and intestinal flora were extracted from the GeneCards and OMIM databases.The in-tersection targets between the drug and disease were identified using a Venn diagram.Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology(GO)enrichment analysis were performed on the intersection targets.Molecular docking technology was em-ployed to validate the binding activities of key components,such as quercetin,kaempferol,with core targets,including TP53 and STAT3.The correlation between targets and the prognosis of liver cancer was assessed through survival analysis.Results KEGG enrichment anal-ysis indicated that shugan formula was primarily enriched in pathways such as cancer pathways.GO enrichment analysis suggested its in-volvement in regulating processes like apoptosis.Molecular docking demonstrated stable binding between key components and core targets.Survival analysis revealed that high expression of tumor protein p53gene(TP53)shortened the overall survival of liver cancer patients,whereas high expression of signal transducer and activator of transcription 3(STAT3)prolonged survival.Conclusion Shugan formula may upregulate STAT3 expression,inhibit TP53 expression,modulate relevant signaling pathways,improve the structure of intestinal flora in liver cancer,and thereby inhibit the proliferation and metastasis of liver cancer cells.

Objective To investigate the renal function recovery and perioperative risk factors for chronic kidney disease in patients after acute Stanford type A aortic dissection (ATAAD) repair. Methods A retrospective study was conducted on patients who underwent ATAAD repair at the Xiamen Cardiovascular Hospital, Xiamen University from 2020 to 2021, and their clinical data were analyzed. Results A total of 255 patients were included, with 200 males and 55 females, and an average age of (52.80±12.46) years. The incidence of acute kidney injury (AKI) after ATAAD repair was 43.9%. Dissection involving the renal artery [OR=2.144, 95%CI (1.234, 3.765), P=0.007], intraoperative urine output [OR=0.761, 95%CI (0.625, 0.911), P=0.004], and intraoperative red blood cell transfusion [OR=1.288, 95%CI (1.088, 1.543), P=0.004] were significantly associated with early AKI after ATAAD repair. Long-term renal function follow-up data were available for 232 patients, among whom 40 (17.2%) patients developed chronic kidney disease (CKD). Independent predictors for CKD included lower body mass index [OR=0.827, 95%CI (0.723, 0.931), P=0.003], preoperative cardiac tamponade [OR=5.344, 95%CI (1.65, 17.958), P=0.005], preoperative renal hypoperfusion syndrome [OR=12.629, 95%CI (5.003, 35.373), P<0.001], postoperative peak serum creatinine time>3 d [OR=7.566, 95%CI (2.799, 22.731), P<0.001], and AKI grade [grade 1: OR=4.418, 95%CI (1.339, 15.361), P=0.016; grade 2: OR=8.345, 95%CI (1.762, 40.499), P=0.007; grade 3: OR=9.463, 95%CI (2.602, 37.693), P<0.001]. Conclusion AKI related to ATAAD repair can recover in the early postoperative period, but both the duration and severity of AKI will affect long-term renal function. In addition, patients' nutritional status, preoperative cardiac tamponade, and renal hypoperfusion syndrome are also independent risk factors for long-term renal dysfunction.

BACKGROUND:Slow bone repair and poor bone formation quality are still problems during masquelet technique in the treatment of large segment bone defects.H-type vessels can induce osteogenesis,enhance the local angiogenesis and osteogenesis coupling,and promote bone repair.However,there are few reports on the role of H-type blood vessels in the bone induced membrane.OBJECTIVE:To construct a large segment bone defect model of SD rat tibia,observe the expression characteristics of H-type blood vessels in the bone induced membrane,then to identify the expression peak point of H-type blood vessels in the bone induced membrane and determine the optimal period of bone grafting.METHODS:Sixty SD rats were randomly divided into a control group(n=30)and a model group(n=30)by random number table method.The two groups were further divided into three subgroups at 4,6,and 8 weeks after bone cement implantation,with 10 rats in each group.A 4 mm bone defect model of the right tibia was constructed in both the control and the model groups.Polymethyl methacrylate bone cement was implanted in the model group to induce bone biomembrane formation,while bone cement was not implanted in the control group.At 4,6,and 8 weeks after bone cement implantation,6 rats were randomly selected at each time point.The bone induction membrane tissue was cut from the model group,and the non-bone soft tissue of the corresponding part was cut from the control group.The dynamic expressions of H-type blood vessels in the bone induced membrane were identified by immunofluorescence.The morphological changes of the bone induced membrane were observed by hematoxylin-eosin staining.The formation of blood vessels in the bone induced membrane was observed by angiography.The expression levels of osteoblast-specific transcription factor in the bone induced membrane were detected by immunohistochemistry.Four rats remained at each time point.In the model group,the bone induced membrane was cut open and the bone cement was removed and autologous coccyx was implanted.In the control group,autologous coccyx was implanted in the bone defect area.Micro-CT evaluation of the tibial defect was performed 8 weeks after bone grafting.RESULTS AND CONCLUSION:(1)Immunofluorescence staining showed that the expression of H-type vessels in the model group was most obvious 6 weeks after bone cement implantation,and the expression of H-type vessels in the model group at each time point after bone cement implantation was higher than that in the control group(P＜0.05).(2)Hematoxylin-eosin staining and angiography showed that the number and volume of new blood vessels at each time point after bone cement implantation in the model group were greater than those in the control group(P＜0.05).The order of the number and volume of new blood vessels in the model group was:8 weeks after bone cement implantation＞6 weeks after bone cement implantation＞4 weeks after bone cement implantation.(3)Immunohistochemical staining showed that the positive expression of osteoblast-specific transcription factors at each time point after bone cement implantation in the model group was higher than that in the control group(P＜0.05),and the positive expression of osteoblast-specific transcription factors in the model group was most obvious 6 weeks after bone cement implantation.(4)Micro-CT detection showed that the bone repair effect of the three subgroups in the model group was significantly better than that of the corresponding subgroups in the control group,and the bone repair effect of the subgroup in the model group 6 weeks after bone cement implantation was better than that of the subgroups 4 and 8 weeks after bone cement implantation.The results indicate that H-type blood vessels are dynamically expressed in the bone induced membrane and reached a peak 6 weeks after bone cement implantation.Good bone repair effects can be obtained by the bone induced membrane bone grafting 6 weeks after bone cement implantation.

Objective:To explore the characteristics and evolutionary features of cognitive impairment and clinical transformation in early-stage Parkinson′s disease (PD) patients.Methods:Based on the cohort of patients with primary unmedicated PD admitted to the Parkinson′s Specialized Outpatient Clinic of Affiliated Brain Hospital of Nanjing Medical University from November 2018 to July 2022, follow-up was conducted for PD patients who completed the baseline assessment and had a follow-up time of 1.5 years or more, and a total of 87 patients finally completed the follow-up and were included in the study. At follow-up, the 87 patients were divided into a cognitively impaired group ( n=36) and a cognitively normal group ( n=51) according to the norm proposed by Professor Jia Jianping and colleagues in 2011 for the Chinese elderly population. Differences in baseline clinical characteristics between the 2 groups were compared, and binary Logistic regression analysis was used to explore risk factors for cognitive impairment in PD patients. In addition, transformed grouping according to cognitive assessment results at baseline and follow-up was used to compare differences in patients′ baseline clinical characteristics among the 3 groups: a reversal group [Parkinson′s disease-mild cognitive impairment (PD-MCI), reverting to Parkinson′s disease-cognitively normal (PD-CN); n=15], a non-reversal group (persistent PD-MCI; n=24), and a stable group (stable PD-CN; n=36). Results:Cognitive reversal occurred at follow-up in 36.6% (15/41) of patients with cognitive impairment at baseline, and 21.7% (10/46) of patients with normal cognition at baseline had cognitive impairment at follow-up. At the end of the follow-up, the 87 patients with PD had higher Unified Parkinson′s Disease Rating Scale Ⅱ (UPDRS-II) scores [8 (6, 11)], Unified Parkinson′s Disease Rating Scale Ⅲ (UPDRS-Ⅲ) scores [23 (16, 30)], and Hoehn-Yahr stages [2.0 (1.5, 2.5)] than those at baseline [7(4, 10), 19(14, 25), 1.5(1.0, 2.0)]. The differences were statistically significant ( Z=-2.498, P=0.012; Z=-3.039, P=0.002; Z=-4.436, P<0.001). The cognitively impaired group had lower Montreal Cognitive Assessment scores [22.00(19.00, 23.75)] and fewer years of education [9.00(8.00, 11.75) years] but higher Parkinson′s Disease Non-Motor Symptoms Questionnaire (PD-NMSQ) scores [8.00(5.25, 12.00)] than the cognitively normal group [25.00(24.00, 27.00), 12.00(9.00, 15.00) years, 6.00(3.00, 8.00)], and the differences were statistically significant ( Z=-4.764, P<0.001; Z=-3.016, P=0.003; Z=-3.281, P=0.001). Multivariate Logistic regression showed that years of education ( OR=0.829, 95%CI 0.715-0.960, P=0.012) and PD-NMSQ scores ( OR=1.200, 95%CI 1.040-1.384, P=0.012) were independent predictors of cognitive impairment in patients with PD. There were statistically significant differences among the reversal, non-reversal, and stable groups in years of education ( F=5.366, P=0.010), PD-NMSQ scores ( H=10.795, P=0.005), and UPDRS-Ⅱ scores ( H=6.957, P=0.031). Pairwise comparisons showed lower PD-NMSQ scores [4.00(3.00, 7.00) vs 8.00(6.25, 12.75); Z=-2.989, P=0.003] and lower UPDRS-Ⅱ scores [6.00(3.00, 6.00) vs 7.00(6.00, 10.00); Z=-2.756, P=0.006] in the reversal group than in the non-reversal group, indicating better baseline quality of life in cognitive reversal patients. Conclusions:Low educational level and severe non-motor symptoms were risk factors predicting cognitive impairment in PD patients. Conversely, mild non-motor symptoms with high quality of life (lower UPDRS-Ⅱ scores) were important factors for cognitive reversal.

The anterior cingulate cortex (ACC) has recently been proposed as a key player in the representation of itch stimuli. However, to date, little is known about the contribution of specific ACC interneuron populations to itch processing. Using c-Fos immunolabeling and in vivo Ca²⁺ imaging, we reported that both histamine and chloroquine stimuli-induced acute itch caused a marked enhancement of vasoactive intestinal peptide (VIP)-expressing interneuron activity in the ACC. Behavioral data indicated that optogenetic and chemogenetic activation of these neurons reduced scratching responses related to histaminergic and non-histaminergic acute itch. Similar neural activity and modulatory role of these neurons were seen in mice with chronic itch induced by contact dermatitis. Together, this study highlights the importance of ACC VIP⁺ neurons in modulating itch-related affect and behavior, which may help us to develop novel mechanism-based strategies to treat refractory chronic itch in the clinic.
Animals ; Pruritus/physiopathology* ; Vasoactive Intestinal Peptide/metabolism* ; Interneurons/metabolism* ; Gyrus Cinguli/metabolism* ; Mice ; Male ; Mice, Inbred C57BL ; Histamine ; Chloroquine ; Optogenetics ; Mice, Transgenic

OBJECTIVES: Esophageal squamous cell carcinoma (ESCC) is characterized by complex pathogenesis and poor prognosis. In recent years, epithelial-mesenchymal transition (EMT) in tumor initiation and progression has attracted increasing attention. Enhancer of zeste homolog 2 (EZH2), which is aberrantly expressed in various tumors, may be closely related to the EMT process. This study aims to examine the expression and correlation of EZH2 and EMT markers in ESCC cells and tissues, evaluate the effects of EZH2 knockdown on ESCC cell proliferation, invasion, and migration, and explore how EZH2 contributes to the malignant biological behavior of ESCC. METHODS: Bioinformatics analyses were used to assess EZH2 expression levels in ESCC. Small interfering RNA was used to knock down EZH2 in ESCC cell lines EC109 and EC9706. Cell proliferation, invasion, and migration were evaluated using cell counting kit-8 (CCK-8), wound healing, and Transwell assays. Protein and mRNA expression levels of EZH2, E-cadherin (E-cad), and vimentin (Vim) were detected by Western blotting and real time fluorogenic quantitative PCR (RT-qPCR), respectively. Immunohistochemical (IHC) staining was performed on 70 ESCC tissue samples and 40 paired adjacent normal tissues collected from the First Affiliated Hospital of Shihezi University between 2010 and 2016 to assess the expression of EZH2, E-cad, and Vim, and to analyze their associations with clinicopathological feature and patient prognosis. RESULTS: Bioinformatics analysis showed that EZH2 was highly expressed in ESCC (P<0.001), and high EZH2 expression was associated with worse prognosis (P<0.001). CCK-8, wound healing, and Transwell assays demonstrated that EZH2 knockdown significantly suppressed the proliferation, invasion, and migration of ESCC cells (P<0.001). In addition, Vim expression was significantly reduced, while E-cad expression was significantly increased at both protein and mRNA levels in EZH2-silenced cells (all P<0.05). IHC staining analysis revealed higher expression of EZH2 and Vim and lower expression of E-cad in ESCC tissues compared to adjacent normal tissues. Kaplan-Meier survival analysis showed that low expression of EZH2 and Vim and high expression of E-cad were associated with longer survival (all P<0.05). CONCLUSIONS EZH2 promotes malignant biological behavior in ESCC by mediating EMT. Elevated EZH2 expression is associated with poor prognosis in ESCC patients.
Humans ; Enhancer of Zeste Homolog 2 Protein/physiology* ; Esophageal Squamous Cell Carcinoma/pathology* ; Epithelial-Mesenchymal Transition/genetics* ; Esophageal Neoplasms/metabolism* ; Cell Proliferation ; Cell Line, Tumor ; Cell Movement ; Cadherins/genetics* ; Vimentin/genetics* ; Male ; Female ; Middle Aged ; Neoplasm Invasiveness ; Prognosis ; RNA, Small Interfering/genetics* ; Gene Expression Regulation, Neoplastic

Ischemic stroke,a major public health challenge,is marked by high incidence,disability,and mortality rates.The pathophysiology of stroke is complex,and inflammation reaction plays a crucial role throughout the process.In recent years,anti-inflammatory treatment has become a major focus in researches of ischemic stroke.Current preclinical and clinical investigations have revealed the potential of various anti-inflammatory strategies for ischemic stroke.However,which patient populations benefit from existing drugs are not yet elucidated.Additionally,drugs targeting innate immune cascades have become a new research hot spot.This article reviewed the latest clinical advances in anti-inflammatory therapy for ischemic stroke and discusses its promising prospects,aiming to provide novel clinical application strategies and future research directions.

BACKGROUND:Slow bone repair and poor bone formation quality are still problems during masquelet technique in the treatment of large segment bone defects.H-type vessels can induce osteogenesis,enhance the local angiogenesis and osteogenesis coupling,and promote bone repair.However,there are few reports on the role of H-type blood vessels in the bone induced membrane.OBJECTIVE:To construct a large segment bone defect model of SD rat tibia,observe the expression characteristics of H-type blood vessels in the bone induced membrane,then to identify the expression peak point of H-type blood vessels in the bone induced membrane and determine the optimal period of bone grafting.METHODS:Sixty SD rats were randomly divided into a control group(n=30)and a model group(n=30)by random number table method.The two groups were further divided into three subgroups at 4,6,and 8 weeks after bone cement implantation,with 10 rats in each group.A 4 mm bone defect model of the right tibia was constructed in both the control and the model groups.Polymethyl methacrylate bone cement was implanted in the model group to induce bone biomembrane formation,while bone cement was not implanted in the control group.At 4,6,and 8 weeks after bone cement implantation,6 rats were randomly selected at each time point.The bone induction membrane tissue was cut from the model group,and the non-bone soft tissue of the corresponding part was cut from the control group.The dynamic expressions of H-type blood vessels in the bone induced membrane were identified by immunofluorescence.The morphological changes of the bone induced membrane were observed by hematoxylin-eosin staining.The formation of blood vessels in the bone induced membrane was observed by angiography.The expression levels of osteoblast-specific transcription factor in the bone induced membrane were detected by immunohistochemistry.Four rats remained at each time point.In the model group,the bone induced membrane was cut open and the bone cement was removed and autologous coccyx was implanted.In the control group,autologous coccyx was implanted in the bone defect area.Micro-CT evaluation of the tibial defect was performed 8 weeks after bone grafting.RESULTS AND CONCLUSION:(1)Immunofluorescence staining showed that the expression of H-type vessels in the model group was most obvious 6 weeks after bone cement implantation,and the expression of H-type vessels in the model group at each time point after bone cement implantation was higher than that in the control group(P＜0.05).(2)Hematoxylin-eosin staining and angiography showed that the number and volume of new blood vessels at each time point after bone cement implantation in the model group were greater than those in the control group(P＜0.05).The order of the number and volume of new blood vessels in the model group was:8 weeks after bone cement implantation＞6 weeks after bone cement implantation＞4 weeks after bone cement implantation.(3)Immunohistochemical staining showed that the positive expression of osteoblast-specific transcription factors at each time point after bone cement implantation in the model group was higher than that in the control group(P＜0.05),and the positive expression of osteoblast-specific transcription factors in the model group was most obvious 6 weeks after bone cement implantation.(4)Micro-CT detection showed that the bone repair effect of the three subgroups in the model group was significantly better than that of the corresponding subgroups in the control group,and the bone repair effect of the subgroup in the model group 6 weeks after bone cement implantation was better than that of the subgroups 4 and 8 weeks after bone cement implantation.The results indicate that H-type blood vessels are dynamically expressed in the bone induced membrane and reached a peak 6 weeks after bone cement implantation.Good bone repair effects can be obtained by the bone induced membrane bone grafting 6 weeks after bone cement implantation.

Objective To elucidate the molecular mechanism of shugan formula in regulating intestinal flora in liver cancer using network pharmacology.Methods The active components and targets of shugan formula were screened using the TCMSP and BATMAN-TCM databases.Targets associated with liver cancer and intestinal flora were extracted from the GeneCards and OMIM databases.The in-tersection targets between the drug and disease were identified using a Venn diagram.Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology(GO)enrichment analysis were performed on the intersection targets.Molecular docking technology was em-ployed to validate the binding activities of key components,such as quercetin,kaempferol,with core targets,including TP53 and STAT3.The correlation between targets and the prognosis of liver cancer was assessed through survival analysis.Results KEGG enrichment anal-ysis indicated that shugan formula was primarily enriched in pathways such as cancer pathways.GO enrichment analysis suggested its in-volvement in regulating processes like apoptosis.Molecular docking demonstrated stable binding between key components and core targets.Survival analysis revealed that high expression of tumor protein p53gene(TP53)shortened the overall survival of liver cancer patients,whereas high expression of signal transducer and activator of transcription 3(STAT3)prolonged survival.Conclusion Shugan formula may upregulate STAT3 expression,inhibit TP53 expression,modulate relevant signaling pathways,improve the structure of intestinal flora in liver cancer,and thereby inhibit the proliferation and metastasis of liver cancer cells.