BACKGROUND:Troxerutin has been found to have a significant ameliorative effect on brain disorders,but there are fewer studies on the effects of troxerutin on the treatment of cerebral infarction and on neuronal cells. OBJECTIVE:To investigate the mechanism by which troxerutin regulates nuclear factor-κB signaling pathway to reduce brain injury and neuronal apoptosis in cerebral infarction rats. METHODS:Fifty clean grade rats were randomized into healthy group,model group,and troxerutin+nuclear factor-κB agonist group,troxerutin group,and nuclear factor-κB inhibitor group.Except for the healthy group,all other groups were used to establish a rat model of cerebral infarction by arterial ligation.The healthy and model groups were treated once a day with an equal amount of physiological saline by gavage.The troxerutin+nuclear factor-κB agonist group was intervened with 72 mg/kg troxerutin by gavage+20 mg/kg RANK intraperitoneally.The troxerutin group was treated with 72 mg/kg troxerutin by gavage.The nuclear factor κB inhibitor group was intervened intraperitoneally with 120 mg/kg nuclear factor κB inhibitor pyrrolidine disulfiram.Administration in each group was given once a day for 30 continuous days.Zea-longa was used to detect neurological damage in rats,hematoxylin-eosin staining was used to observe pathological changes,TUNEL was used to detect neuronal apoptosis,and immunoblotting and PCR were used to detect the expression of nuclear factor-κB p65 and nuclear factor-κB p50 at protein and mRNA levels,respectively. RESULTS AND CONCLUSION:Compared with the healthy group,the neurological function score,neuronal apoptosis rate,nuclear factor-κB p65,nuclear factor-κB p50 mRNA and protein expression levels were elevated in the model group(P＜0.05).Compared with the model group,the neurological function score,neuronal apoptosis rate,nuclear factor-κB p65 and nuclear factor-κB p50 mRNA and protein expression levels were decreased in the troxerutin+nuclear factor-κB agonist group(P＜0.05).Compared with the troxerutin+nuclear factor-κB agonist group,the neurological function score,neuronal apoptosis rate,nuclear factor-κB p65 and nuclear factor-κB p50 mRNA and protein expression levels were reduced in the troxerutin group and nuclear factor-κB inhibitor group(P＜0.05).In addition,there was no difference between the troxerutin group and the nuclear factor-κB inhibitor group(P＞0.05).In the model group,there was a large number of cytoplasmic vacuolation,obvious edema and necrosis,and a large number of inflammatory cell infiltrations.In the troxerutin+nuclear factor-κB agonist,the swelling of brain tissue was reduced,and reticulate structures and condensed cells were reduced,still with some edema.In the troxerutin group and nuclear factor-κB inhibitor group,brain tissue swelling,neuronal edema degeneration,cytoplasmic vacuolation and neuronal nucleus consolidation were reduced,and the inflammatory cell infiltration was significantly decreased.To conclude,troxrutin can reduce the expression of neurological impairment,inhibit neuronal apoptosis and improve the pathological injury of brain tissue in rats with cerebral infarction,and its mechanism of action may be related to the modulation of nuclear factor-κB expression and related signaling pathways.

ObjectiveTo investigate the regulatory effects of Ginkgolide B on the biological characteristics of brain T cells and their interactions with glial cells during the recovery phase of ischemic stroke in mice. Methods36 adult C57BL/6 mice were randomly assigned to three groups: sham-operated group (Sham group), control group (PBS group), and Ginkgolide B treatment group (GB group). The Sham group underwent only sham surgeries, whereas the PBS and GB groups were subjected to a middle cerebral artery occlusion (MCAO) model using the filament method, followed by intranasal administration of an equivalent volume of either PBS or Ginkgolide B solution for 14 days post-injury. Neurological function changes were evaluated in all three groups using the rotarod test and a neurological scoring system. On day 15, single-cell sequencing was performed on fresh tissues from the brain injury areas, surrounding cortex, corpus callosum, and striatum of mice in the PBS and GB group to assess the biological characteristics of T cells and their subpopulations, and further explore the interactions and mechanisms among T cells, microglia, and oligodendrocytes. ResultsCompared with the Sham group, both PBS and GB group exhibited significant improvements in neurological scores and reduced pre-fall motor durations (P < 0.001). Compared with the PBS group, the GB group showed a downward trend in neurological scores and an upward trend in pre-fall motor durations on days 5, 10, and 15 post-ischemic brain injury, with a significant increase in pre-fall motor duration on day 15 (P < 0.05). Compared with the PBS group, the GB group exhibited a significant increase in T cell proliferative activity in the brain 15 days post brain injury (P < 0.05). The number of proliferative T cells and the levels of lipid metabolism were significantly elevated (P < 0.05), and there was a significant increase in extracellular matrix remodeling in all T cells (P < 0.05). Additionally, the interactions between T cells and both microglia and oligodendrocytes, as well as among the microglia themselves and between microglia and oligodendrocytes, were significantly enhanced in the GB group. This was primarily evident in the strengthened interactions between CD74 and macrophage migration inhibitory factor (MIF), as well as colony stimulating factor 1 receptor (CSF1R) and colony stimulating factor 1 (CSF1) (P < 0.05). However, the inflammatory levels of T cells showed no significant differences compared with the PBS group. ConclusionA mouse model of ischemic stroke can be successfully established by MCAO operation. Ginkgolide B may promote neurological recovery post-brain injury in mice by modulating the biological characteristics of T cells within the brain and their interactions with glial cells.

Objective:To investigate the effects of the taurochenodeoxycholic acid(TCDCA)on cartilage degeneration in tem-poromandibular joint osteoarthritis(TMJ OA).Methods:36 female SD rats aged 6 weeks were randomly divided into 3 groups:con-trol group(CON),unilateral anterior crossbite group(UAC),UAC plus TCDCA injection group(UAC+TCDCA).UAC model was es-tablished in all rats in UAC and UAC+TCDCA groups.Samples were collected at 8 and 12 weeks(control group,UAC group,UAC+TCDCA group)after set up of the experiment(n=6),and TMJ morphological examination was performed.The expression of CYP7A1,BAAT and TGR5 in the tissue and cells was examined by immunohistochimical staining.Results:(1)Compared with the CON group of the same age,the cells in the condylar cartilage were disordered,the cartilage matrix was reduced and thinner in UAC group.Compared with UAC group of the same age,cell arrangement,cell number,cartilage matrix and cartilage thickness were im-proved in UAC+TCDCA group(P＜0.05).(2)Compared with the CON group of the same age,the positive cells for TCDCA-specific receptor TGR5 and the key enzymes CYP7A1 and BAAT were mainly distributed in the anterior hypertrophic layer and hypertrophic layer at each time point.The number of positive cells in the UAC group was significantly reduced compared with the CON group.Conclusion:TCDCA has obvious therapeutic effects on the degeneration in TMJ OA.

Objective To investigate the feasibility, safety, and short-term efficacy of minimally invasive McKeown esophagectomy (MIME) in patients with locally advanced thoracic esophageal squamous cell carcinoma (TESCC) after neoadjuvant immunotherapy. Methods The clinical data of the patients with locally advanced TESCC in the First Affiliated Hospital of University of Science and Technology of China from July 2022 to March 2023 were restrospectively analyzed. They were divided into a neoadjuvant immunotherapy (NI) group and a non-neoadjuvant immunotherapy (NNI) group according to different preoperative neoadjuvant therapy. The perioperative clinical data and 3-month follow-up data were compared between the two groups. Results A total of 47 patients were collected, including 31 males and 16 females with a mean age of (67.57±7.64) years. There were 29 patients in the NI group and 18 patients in the NNI group. There were no statistical differences in baseline data, perioperative complications, short-term complications, surgical time, intraoperative bleeding, postoperative adjuvant therapy, metastasis/recurrence within 3 months, R0 resection rate, postoperative pathological staging decline, or College of American Pathologists (CAP) tumor regression grade between the two groups (P＞0.05). Conclusion Neoadjuvant immunotherapy combined with minimally invasive McKeown esophagectomy can be safely and effectively performed for patients with locally advanced TESCC without increasing operation time, intraoperative blood loss and perioperative complications.

T cell immunoglobulin and ITIM domain (TIGIT) is a novel immune checkpoint that has been considered as a target in cancer immunotherapy. Current available bioassays for measuring the biological activity of therapeutic antibodies targeting TIGIT are restricted to mechanistic investigations because donor primary T cells are highly variable. Here, we designed a reporter gene assay comprising two cell lines, namely, CHO-CD112-CD3 scFv, which stably expresses CD112 (PVRL2, nectin-2) and a membrane-bound anti-CD3 single-chain fragment variable (scFv) as the target cell, and Jurkat-NFAT-TIGIT, which stably expresses TIGIT as well as the nuclear factor of activated T-cells (NFAT) response element-controlled luciferase gene, as the effector cell. The anti-CD3 scFv situated on the target cells activates Jurkat-NFAT-TIGIT cells through binding and crosslinking CD3 molecules of the effector cell, whereas interactions between CD112 and TIGIT prevent activation. The presence of anti-TIGIT mAbs disrupts their interaction, which in turn reverses the inactivation and luciferase expression. Optimization and validation studies have demonstrated that this assay is superior in terms of specificity, accuracy, linearity, and precision. In summary, this reliable and effective reporter gene assay may potentially be utilized in lot release control, stability assays, screening, and development of novel TIGIT-targeted therapeutic antibodies.

The use of traditional finite element method (FEM) in occlusal stress analysis is limited due to the complexity of musculature simulation. The present purpose was to develop a displacement boundary condition (DBC)-FEM, which evaded the muscle factor, to predict the dynamic occlusal stress. The geometry of the DBC-FEM was developed based on the scanned plastic casts obtained from a volunteer. The electrognathographic and video recorded jaw positional messages were adopted to analyze the dynamic occlusal stress. The volunteer exhibited asymmetrical lateral movements, so that the occlusal stress was further analyzed by using the parameters obtained from the right-side eccentric movement, which was 6.9 mm long, in the stress task of the left-side eccentric movement, which was 4.1 mm long. Further, virtual occlusion modification was performed by using the carving tool software aiming to improve the occlusal morphology at the loading sites. T-Scan Occlusal System was used as a control of the in vivo detection for the location and strength of the occlusal contacts. Data obtained from the calculation using the present developed DBC-FEM indicated that the stress distribution on the dental surface changed dynamically with the occlusal contacts. Consistent with the T-Scan recordings, the right-side molars always showed contacts and higher levels of stress. Replacing the left-side eccentric movement trace by the right-side one enhanced the simulated stress on the right-side molars while modification of the right-side molars reduced the simulated stress. The present DBC-FEM offers a creative approach for pragmatic occlusion stress prediction.

Objective:To describe and analyze the clinical manifestations of patients with orofacial pain of temporomandibular disorders (TMD).Methods:A retrospective study on orofacial pain was conducted for 3 425 patients diagnosed as TMD based on clinical symptoms and signs in the Department of Temporomandibular Disorders and Orofacial Pain, School of Stomatology, The Fourth Military Medical University. The patients included 1 158 males and 2 267 females with a median age of 32 years. The gender, age, course of disorders, pattern and site of pain, CT imaging diagnosis of temporomandibular joint (TMJ) were analyzed. The distribution of gender, age and disorder course interval were described. The differences in frequency of the pattern and site of pain, imaging diagnosis in different gender, age and disease course interval were compared. Chi-square test and non-parametric rank sum test were performed using software SPSS 23.0.Results:Of the 3 425 patients, 29.1% (997/3 245) had signs of joint popping, and 40.1% (1 373/3 425) had restricted opening. The pain frequency was higher in males who had disorder course less than 1 month ( P<0.01) and also in males who had open-and-close and/or lateral excursion and/or protrusion pain without tenderness or other pain without tenderness ( P<0.05). However, the pain frequency was higher in females who had tenderness ( P<0.01). The pain frequencies in those over 56 years old with tenderness combined with open-and-close and/or lateral excursion and/or protrusion pain were higher than in patients of other ages ( P<0.01). In patients with unilateral TMJ pain, the frequency in males was higher than females( P<0.01), while the frequency in females was higher in patients with unilateral TMJ pain combined with unilateral or bilateral myalgia and the frequency was higher in patients under 15 years old having bilateral TMJ pain and/or unilateral or bilateral myalgia ( P<0.05). In patients with unilateral TMJ pain, the frequency in those with disorder course≤1 month was higher than in those with other disease duration intervals ( P<0.01), while in patients with bilateral myalgia or TMJ pain plus unilateral or bilateral myalgia, the frequency in those with disorder course>3 years was higher than in those with other disease duration intervals( P<0.01). In patients with unilateral TMJ pain, the frequency was higher in those having open-and-close and/or lateral excursion and/or protrusion pain ( P<0.01). In patients with unilateral myalgia and bilateral myalgia, the frequency was higher in those having tenderness ( P<0.01). The frequency of TMJ space changes in male patients was higher than females and the frequency of hyperosteogeny and resorption in females were higher than males ( P<0.05). The frequency of TMJ space changes and developmental problems were higher in patients aged 16 to 35 years, while the frequencies of hyperosteogeny, bone resorption and cystis in those over 56 years were higher than other ages ( P<0.01). The frequency of TMJ space changes in patients with disorder course≤1 month was higher than in those with other disease duration intervals ( P<0.01), while the frequency of hyperosteogeny was higher in patients with disorder course>3 years ( P<0.01). Conclusions:The male to female ratio in the present patients with orofacial pain of TMD was about 1 to 2. Most of the patients visited hospital within half a year after the disorders occurred. The pattern and site of the orofacial pain, signs on TMJ CT images showed some distribution regularities in views of gender, age and disorder course.

More efficient drug delivery system and formulation with less adverse effects are needed for the clinical application of broad-spectrum antineoplastic agent doxorubicin (DOX). Here we obtained outer-membrane vesicles (OMVs), a nano-sized proteoliposomes naturally released by Gram-negative bacteria, from attenuated and prepared doxorubicin-loaded O0MVs (DOX-OMV). Confocal microscopy and distribution study observed that DOX encapsulated in OMVs was efficiently transported into NSCLC A549 cells. DOX-OMV resulted in intensive cytotoxic effects and cell apoptosis as evident from MTT assay, Western blotting and flow cytometry due to the rapid cellular uptake of DOX. In A549 tumor-bearing BALB/c nude mice, DOX-OMV presented a substantial tumor growth inhibition with favorable tolerability and pharmacokinetic profile, and TUNEL assay and H&E staining displayed extensive apoptotic cells and necrosis in tumor tissues. More importantly, OMVs' appropriate immunogenicity enabled the recruitment of macrophages in tumor microenvironment which might synergize with their cargo DOX . Our results suggest that OMVs can not only function as biological nanocarriers for chemotherapeutic agents but also elicit suitable immune responses, thus having a great potential for the tumor chemoimmunotherapy.

Objectives: To evaluate the diagnostic value of CT for lymph node metastasis of thoracic esophageal carcinoma with a diameter of more than 1 cm, and to find the optimal diagnostic index by comparing relevant CT indexes. Methods: Totally 80 patients with pathologically proved thoracic esophageal cancer with preoperative CT examination revealed lymph node diameter greater than 1 cm admitted at Department of Thoracic Surgery, the First Affiliated Hospital of University of Science and Technology of China from January 2016 to January 2018 were enrolled in this study. There were 70 males and 10 females, aging of (60±14) years (range: 40-85 years). According to the pathological result of lymph nodes, all the patients and lymph nodes were divided into two groups (N+group: 47 patients, 62 lymph nodes; N-group: 33 patients, 39 lymph nodes). The average number of dissected lymph nodes were 21±4 and 101 lymph nodes′ diameter were greater than 1 cm. The clinicopathologic factors, postoperative complications, lymph node dissection and relevant CT indexes like the minimum diameter of lymph nodes (Min D), the maximum diameter of lymph node (Max D), lymph node axial ratio(LAR), the enhancement of lymph node (ELN) and the boundary of lymph node (BLN) were compared. The clinicopathological data, lymph node dessection and CT parameters of the two groups were compared by t test, χ² test or Wilcoxon rank sum test. Receiver operating characteristic (ROC) curve analysis was used to compare the ability to predict lymph node metastasis between Min D, Max D, LAR, ELN and BLN. Multiple Logistic regression analysis were performed to determine the independent variables for prediction of lymph node metastasis. Results: The difference of tumor segmentation, pN stage, pTNM stage, total number of metastatic lymph nodes, total number of abdominal lymph node metastases, Min D, Max D, ELN and BLN between the two groups were statistically significant. The results of univariate and multivariate analyses showed that gender (OR=0.128, 95%CI: 0.019 to 0.858, P=0.034), pTNM stage (OR=1.514, 95%CI: 1.020 to 2.247, P=0.039), Min D (OR=0.102, 95%CI: 0.010 to 0.995, P=0.050) and LAR (OR=0.195, 95%CI: 0.052 to 0.731, P=0.015) were the independent relative factors. The area under the curve of ROC curve analysis of Min D, Max D, LAR, ELN and BLN were 0.679, 0.666, 0.561, 0.650 and 0.820, respectively. BLN was the best CT index to diagnosis lymph node metastasis, while the accuracy of dignosis of lymph node metastasis of BLN was 97.0%. The Youden index of Min D, Max D and LAR were 1.25 cm, 1.64 cm and 0.77, respectively. Combining the BLN and ELN had a higher diagnostic rate (97.0%) of lymph node metastasis. Conclusions CT has high diagnostic value for lymph node metastasis in thoracic esophagel cancer when the lymph node diameter is greater than 1 cm. BLN is the best diagnostic index for lymph node metastasis.

To investigate the feasibility, safety and short-term efficacy of modified inflatable video-mediastinoscopy in patients with early esophageal cancer. Methods The study retrospectively evaluated 54 patients with cT1N0M0 esophageal carcinoma who received minimally invasive esophagectomy in the First Affiliated Hospital of University of Science and Technology of China between July 2017 and June 2018. Of those patients, 23 patients underwent modified inflatable video-assisted mediastinoscopic transhiatal esophagectomy(MIVMTS) and 31 underwent minimally invasive McKeown esophagectomy (MIME). The clinicopathologic factors, operational factors, postoperative complications and lymph node dissection of patients were compared. Results There was no significant difference in clinicopathological data between the MIVMTS group and MIME group. The incidence of total minor postoperative complications, pulmonary infection of minior postoperative complications, total postoperative complications and total pulmonary complications in MIME group were higher than MIVMTS group. The incidence of recurrent laryngeal nerve injury, arrhythmia and air leaks in minior and pulmonary infection, chylothorax, anastomotic fistula in major postoprative complications were no different in the two groups with P >0. 05. The intraoperative blood loss, duration of surgery and postoperative thoracic drainage fluid volume of MIVMTS group were less than MIME group, the difference was statistically significant. The postoperative hospitalization of the two groups have no statistics significance(P >0. 05). There were no significant difference in the lymph node dessection of the left laryngeal recurrent nerve lymph nodes, paraesophageal lymph nodes, subcarinal lymph nodes and superior phrenic lymph node of the two groups. However, when compared with MIVMTS group, the MIME group have advantage in the right laryngeal recurrent para-nerve lymph node dissection. Conclusion MIVMTS can be safely and effectively performed for early esophageal cancer with favorable short-term efficacy.