ObjectiveTo investigate the mechanisms of Tongnao decoction （TND） in mice with acute ischemic stroke （AIS）. MethodsFifty male C57BL/6J mice were randomly divided into a sham operation group， model group， TND low-dose group （1.86 g·kg^-1）， TND high-dose group （3.72 g·kg^-1）， and butylphthalide （NBP） group （10 mg·kg^-1）， with 10 mice in each group. A mouse model of cerebral ischemic injury was established using photochemical thrombosis （PT）. The sham operation group and model group were administered an equal volume of normal saline by gavage. All five groups were treated once daily for 14 consecutive days. Behavioral tests were performed before modeling and at the end of administration. T₂-weighted imaging （T₂WI） was performed 3 days after modeling to evaluate the extent of injury. Hematoxylin-eosin （HE） staining was used to observe histological changes in the cerebral cortex， and Nissl staining was used to observe neuronal morphology. Cerebral blood flow in mice was detected using a laser speckle contrast imaging （LSCI） system. Immunofluorescence staining was used to detect the cell proliferation marker bromodeoxyuridine （BrdU） and the highly glycosylated type I transmembrane glycoprotein CD34. Western blot analysis was used to detect the expression levels of phosphatidylinositol 3-kinase （PI3K）， protein kinase B （Akt）， glycogen synthase kinase-3β （GSK-3β）， and their phosphorylation levels， as well as tight junction-related proteins zonula occludens-1 （ZO-1）， Occludin， and Claudin-5 in the peri-infarct tissue. Thirty-five zebrafish were randomly divided into normal control group， model group， TND low and high dose groups （0.16， 0.32 g·L^-1） and NBP group （10 μmol·L^-1）， with 7 in each group. A stereoscopic fluorescence microscope was used to observe vascular growth in zebrafish. ResultsImaging showed that PT caused ischemia in the right cortical region. Behavioral tests indicated that， compared with the model group， the drug-treated groups reduced the error rate of irregular balance ladder climbing on the affected side and shortened the tape removal time （P<0.05）. HE staining and Nissl staining showed that， compared with the model group， the drug-treated groups exhibited reduced brain tissue damage， fewer scars， and improved neuronal morphology. LSCI results showed that the drug-treated groups partially restored cerebral blood perfusion and promoted the establishment of collateral circulation compared with the model group. Immunofluorescence staining indicated that the drug-treated groups increased the positive rates of BrdU and CD34 compared with the model group （P<0.01）， promoting angiogenesis. Meanwhile， compared with the model group， the drug-treated groups upregulated the expression levels of p-PI3K， p-Akt， p-GSK-3β， and tight junction proteins ZO-1， Occludin， and Claudin-5 （P<0.05，P<0.01）， and increased the number of intersegmental vessels in zebrafish （P<0.05，P<0.01）. ConclusionTND can promote angiogenesis around the infarct in PT model mice by regulating the PI3K/Akt/GSK-3β signaling pathway， thereby improving cerebral ischemic injury.

Objective To investigate the effect of Porphyromonas gingivalis(P.gingivalis)on Th17/Treg imbalance and vascular endothelial function in ApoE-/-mice.Methods Eight-week-old ApoE-/-mice were inoculated orally with P.gingivalis every other day and received high-fat diet for 8 weeks.The alveolar bone loss was assessed by Micro-CT and the oral microflora of mice was analyzed by 16S rRNA sequencing technology.HE staining and oil red O staining were used to detect plaque formation in ApoE-/-mice.The total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C)levels of mice were detected by biochemical detection technology.Real-time quantitative PCR(qRT-PCR)and immunohistochemistry(IHC)were used to detect the indicators of endothelial injury in the model aortic plaques,including vascular cell adhesion molecule(VCAM),intercellular adhesion molecule(ICAM),matrix metallopeptidase(MMP)-2 and MMP-9.The relative proportions of Th17 and Treg in aortic plaque and spleen were detected by IHC,qRT-PCR and flow cytometry,including interleukin-17(IL-17),retinoic acid related orphan receptor γt(RORγt)and forkhead box P3(Foxp3).Results In ApoE-/-mice infected with P.gingivalis,the severity of alveolar bone loss was increased(P＜0.01);the diversity of oral microbiota was decreased(P＜0.05),and the composition of oral microbiota was changed.The proportion of atherosclerotic plaque area was increased(P＜0.01),paralleled by elevated serum levels of TG,TC and LDL-C(P＜0.05),alongside a significant de-crease in HDL-C(P＜0.01).Aortic plaques exhibited upregulated protein expression of VCAM and ICAM as well as heightened mRNA levels of ICAM,VCAM,MMP-2 and MMP-9(P＜0.05).The levels of IL-17 related protein and RORγt transcriptional activity in aortic plaque were increased(P＜0.01),while the levels of Foxp3 related protein and gene were decreased(P＜0.01).The ratio of Th17/Treg cells in spleen was imbalanced(P＜0.05).Conclusion Periodontitis caused by the imbalance of the oral flora of ApoE-/-mice induced by P.gingivalis can cause Th17/Treg imbalance in aortic plaque and exacerbate the endothelial dysfunction during ather-osclerosis.

Objective To investigate the effect of Porphyromonas gingivalis(P.gingivalis)on Th17/Treg imbalance and vascular endothelial function in ApoE-/-mice.Methods Eight-week-old ApoE-/-mice were inoculated orally with P.gingivalis every other day and received high-fat diet for 8 weeks.The alveolar bone loss was assessed by Micro-CT and the oral microflora of mice was analyzed by 16S rRNA sequencing technology.HE staining and oil red O staining were used to detect plaque formation in ApoE-/-mice.The total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C)levels of mice were detected by biochemical detection technology.Real-time quantitative PCR(qRT-PCR)and immunohistochemistry(IHC)were used to detect the indicators of endothelial injury in the model aortic plaques,including vascular cell adhesion molecule(VCAM),intercellular adhesion molecule(ICAM),matrix metallopeptidase(MMP)-2 and MMP-9.The relative proportions of Th17 and Treg in aortic plaque and spleen were detected by IHC,qRT-PCR and flow cytometry,including interleukin-17(IL-17),retinoic acid related orphan receptor γt(RORγt)and forkhead box P3(Foxp3).Results In ApoE-/-mice infected with P.gingivalis,the severity of alveolar bone loss was increased(P＜0.01);the diversity of oral microbiota was decreased(P＜0.05),and the composition of oral microbiota was changed.The proportion of atherosclerotic plaque area was increased(P＜0.01),paralleled by elevated serum levels of TG,TC and LDL-C(P＜0.05),alongside a significant de-crease in HDL-C(P＜0.01).Aortic plaques exhibited upregulated protein expression of VCAM and ICAM as well as heightened mRNA levels of ICAM,VCAM,MMP-2 and MMP-9(P＜0.05).The levels of IL-17 related protein and RORγt transcriptional activity in aortic plaque were increased(P＜0.01),while the levels of Foxp3 related protein and gene were decreased(P＜0.01).The ratio of Th17/Treg cells in spleen was imbalanced(P＜0.05).Conclusion Periodontitis caused by the imbalance of the oral flora of ApoE-/-mice induced by P.gingivalis can cause Th17/Treg imbalance in aortic plaque and exacerbate the endothelial dysfunction during ather-osclerosis.

Objective:To evaluate the cognitive status for oral genetics among graduate students of stomatology, and to provide advice for teaching reform and design of oral genetics course.Methods:A questionnaire survey was conducted among first-year graduate students of stomatology in Medical School of Nanjing University enrolled in the academic year of 2022-2023. The questionnaire was designed for "Assessing cognition and attitudes towards oral genetics" and included four parts: assessment of students' level of awareness and knowledge about oral genetic disorders; understanding the role of genetic factors in stomatology; the causes of oral diseases; and awareness of oral genetic disorders and rare oral diseases. Students completed the questionnaire anonymously and voluntarily at the beginning and end of the course. Excel 2013 was used for data processing. SPSS 21.0 was used for rank sum test, chi-square test, and t-test for paired samples. Results:A total of 58 questionnaires were recovered (recovery rate 89.66%) and 52 questionnaires were valid (26 each at the beginning and end of the course). A total of 26 students aged (24.73±1.28) years were enrolled and 69.23% of the students treated patients with oral genetic disorders. Their knowledge of oral genetics increased significantly after the course ( Z=-3.74, P<0.001). Their familiarity with the number of rare oral diseases and oral genetic disorders increased significantly after the course [(18.34±8.61) vs. (29.69±10.39), t=5.95, P<0.001]. Conclusions:Students generally realize the importance of genetics in stomatology. The implementation of the course helps to improve students' cognition and clinical diagnosis and treatment of oral genetic diseases.

ObjectiveTo evaluate the effects of Tongnaoyin on the blood-brain barrier status and neurological impairment in acute ischemic stroke （AIS） patients with the syndrome of phlegm-stasis blocking collaterals by dynamic contrast-enhanced magnetic resonance imaging （DCE-MRI）. MethodsA total of 63 patients diagnosed with AIS in the Jiangsu Province Hospital of Chinese Medicine from October 2022 to December 2023 were enrolled in this study. According to random number table method，the patients were assigned into a control group （32 cases） and an observation group （31 cases）. The control group received conventional Western medical treatment，and the observation group took 200 mL Tongnaoyin after meals，twice a day from day 2 of admission on the basis of the treatment in the control group. After 7 days of treatment，the patients were examined by DCE-MRI. The baseline data for two groups of patients before treatment were compared. The National Institute of Health Stroke Scale （NIHSS） score and modified Rankin Scale （mRS） score were recorded before treatment and after 90 days of treatment for both groups. The rKtrans，rKep，and rVe values were obtained from the region of interest （ROI） of the infarct zone/mirror area and compared between the two groups. ResultsThere was no significant difference in the NIHSS or mRS score between the two groups before treatment. After 90 days of treatment，the NIHSS and mRS scores declined in both groups，and the observation group had lower scores than the control group （P<0.05）. After treatment，the rKtrans and rVe in the observation group were lower than those in the control group （P<0.01）. ConclusionCompared with conventional Western medical treatment alone，conventional Western medical treatment combined with Tongnaoyin accelerates the repair of the blood-brain barrier in AIS patients，thereby ameliorating neurological impairment after AIS to improve the prognosis.

During the progression of ischemic stroke,cells experience oxidative stress and intracellular energy depletion,triggering the dynamic assembly and disassembly of stress granules.Stress granules may have a dual role in ischemic stroke.In the ultra-early reperfusion period(6-36 h),under acute stress,the stress granules may inhibit neuronal apoptosis and exert neuroprotective effects through reversible liquid-liquid phase separation.In contrast,in the acute reperfusion period(36 h to 2 weeks),under prolonged stress,pathological stress granules may accumulate through liquid-solid phase transition,leading to neuronal dysfunction and inducing ischemic stroke sequelae such as motor and cognitive impairments.This article reviewed the mechanisms of stress granules dynamic phase transitions during ischemic stroke and their effects on neurons,aiming to provide references and insights for future stress granule-targeted interventions for ischemic stroke and its sequelae.

During the progression of ischemic stroke,cells experience oxidative stress and intracellular energy depletion,triggering the dynamic assembly and disassembly of stress granules.Stress granules may have a dual role in ischemic stroke.In the ultra-early reperfusion period(6-36 h),under acute stress,the stress granules may inhibit neuronal apoptosis and exert neuroprotective effects through reversible liquid-liquid phase separation.In contrast,in the acute reperfusion period(36 h to 2 weeks),under prolonged stress,pathological stress granules may accumulate through liquid-solid phase transition,leading to neuronal dysfunction and inducing ischemic stroke sequelae such as motor and cognitive impairments.This article reviewed the mechanisms of stress granules dynamic phase transitions during ischemic stroke and their effects on neurons,aiming to provide references and insights for future stress granule-targeted interventions for ischemic stroke and its sequelae.

Objective:To evaluate the cognitive status for oral genetics among graduate students of stomatology, and to provide advice for teaching reform and design of oral genetics course.Methods:A questionnaire survey was conducted among first-year graduate students of stomatology in Medical School of Nanjing University enrolled in the academic year of 2022-2023. The questionnaire was designed for "Assessing cognition and attitudes towards oral genetics" and included four parts: assessment of students' level of awareness and knowledge about oral genetic disorders; understanding the role of genetic factors in stomatology; the causes of oral diseases; and awareness of oral genetic disorders and rare oral diseases. Students completed the questionnaire anonymously and voluntarily at the beginning and end of the course. Excel 2013 was used for data processing. SPSS 21.0 was used for rank sum test, chi-square test, and t-test for paired samples. Results:A total of 58 questionnaires were recovered (recovery rate 89.66%) and 52 questionnaires were valid (26 each at the beginning and end of the course). A total of 26 students aged (24.73±1.28) years were enrolled and 69.23% of the students treated patients with oral genetic disorders. Their knowledge of oral genetics increased significantly after the course ( Z=-3.74, P<0.001). Their familiarity with the number of rare oral diseases and oral genetic disorders increased significantly after the course [(18.34±8.61) vs. (29.69±10.39), t=5.95, P<0.001]. Conclusions:Students generally realize the importance of genetics in stomatology. The implementation of the course helps to improve students' cognition and clinical diagnosis and treatment of oral genetic diseases.

ObjectiveTo investigate the changes in cerebral blood perfusion in patients with acute cerebral infarction after taking Tongnaoyin， a traditional Chinese medicine， based on head and neck computed tomography （CT） angiography （CTA） combined with brain CT perfusion imaging （CTP）. MethodA total of 240 patients with cerebral infarction of phlegm and blood stasis syndrome treated in Jiangsu Province Hospital of Traditional Chinese Medicine from March 2018 to September 2023 were randomly divided into a control group （99 cases） and a Tongnaoyin group （141 cases）. Based on the guidelines， the control group was treated with conventional treatment such as anti-aggregation， anticoagulation， lipid-lowering and plaque stabilization， brain protection， and supportive treatment. The Tongnaoyin group was treated with Tongnaoyin of 200 mL in warm conditions in the morning and evening on the basis of the control group. Both groups underwent CTA combined with CTP within 24 hours after admission， and they were reexamined by CTA and CTP in the sixth month after admission. The degree of intracranial artery stenosis was determined according to the North American Symptomatic Carotid Endarterectomy Trial （NASCET） method. The relative cerebral blood volume （rCBV）， relative cerebral blood flow （rCBF）， mean transit time （MTT）， and time to peak （TTP） of the lesion area before and after treatment were compared. The adverse outcomes of the two groups within six months after discharge were compared. ResultCompared with the group before treatment， the degree of vascular stenosis in the Tongnaoyin group was significantly reduced， and the difference was statistically significant （Z=105.369，P<0.05）. Compared with the control group after treatment， the improvement rate of vascular stenosis in the Tongnaoyin group was higher （χ²=84.179，P<0.01）， and the curative effect was better.After treatment， the rCBV and rCBF of patients in the Tongnaoyin group were significantly increased， and the difference was statistically significant （P<0.01）. MTT and TTP showed a trend of shortening， but the difference was not statistically significant. There was no statistically significant difference in rCBV， rCBF， MTT， and TTP in the control group. Compared with those in the control group after treatment， the rCBV and rCBF in the Tongnaoyin group were significantly increased， while MTT and TTP were significantly reduced （P<0.01）. After six months of discharge， the risk of poor prognosis in the Tongnaoyin group was significantly reduced compared with the control group （P<0.05）. ConclusionTongnaoyin has a good effect on improving cerebral blood perfusion in patients with acute cerebral infarction. It can be used as an effective supplement for the conventional treatment of ischemic stroke to improve clinical efficacy.

As research delves deeper into the mechanisms of tumor immune responses,studies reveal the importance of microbial com-munities within the tumor microenvironment in tumor progression and their interactions with the host immune system.Intratumoral micro-biota could influence the tumor microenvironment,thereby promoting or inhibiting tumor growth and development.Despite this import-ance,the specific role of intratumoral microbiota impacting cancer immunotherapeutic efficacy remains largely unexplored.A deeper under-standing of the characteristics and biological functions of tumor-specific microbiota heralds a potential revolutionary innovation in cancer treatment.In this review,we introduce the discovery and sources of intratumoral microbiota,also addressing its composition,and discuss tumor tissue characteristics.Moreover,we briefly review the history of cancer immunotherapy development with a particular focus on the research progress concerning the impact of intratumoral microbiota on cancer immunotherapy.Furthermore,we explore emerging strategies that combine targeting intratumoral microbiota with immunotherapy to enhance immune efficacy,inhibit tumor progression,and improve cure rates,anticipating that this approach could represent a new direction for enhancing treatment outcomes and prospects.