ObjectiveTo elucidate the chemical composition of the reference sample of Jinshui Liujunjian and its distribution characteristics in blood and tissues of rats. MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry（UPLC-Q-TOF-MS/MS） was used to detect the reference sample solution， plasma， and tissue samples of Jinshui Liujunjian under positive and negative ion modes， respectively. Qualitative Analysis 10.0 software and a self-constructed database were employed for primary mass spectrum matching.Compound identification was further validated by comparing retention times， secondary mass spectral fragments， reference standards， and literature data to deduce fragmentation pathways. ResultsA total of 122 compounds were identified in the reference sample of Jinshui Liujunjian， including 47 flavonoids， 5 amino acids， 13 iridoids， 16 triterpenoid saponins， etc.， of which 42 compounds were confirmed by comparison with reference substances. A total of 21 prototype components were identified in blood components； 50 prototype components were identified in different tissues， among which 13， 10， 7， 21， 11， 6， 14， and 40 prototype components were identified in the heart， liver， spleen， lung， kidney， brain， large intestine， and stomach， respectively. Among them， 7 compounds such as ferulic acid， glycyrrhizic acid， and nobiletin were exposed in the target organs of lung and kidney. ConclusionThis study elucidates the material basis of the reference samples of Jinshui Liujunjian， primarily composed of flavonoids and triterpenoid saponins， along with their in vivo distribution characteristics. These findings provide a scientific basis for establishing quality evaluation indicators and offer references for subsequent pharmacodynamic and pharmacokinetic investigations.

ObjectiveTo elucidate the chemical composition of the reference sample of Jinshui Liujunjian and its distribution characteristics in blood and tissues of rats. MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry（UPLC-Q-TOF-MS/MS） was used to detect the reference sample solution， plasma， and tissue samples of Jinshui Liujunjian under positive and negative ion modes， respectively. Qualitative Analysis 10.0 software and a self-constructed database were employed for primary mass spectrum matching.Compound identification was further validated by comparing retention times， secondary mass spectral fragments， reference standards， and literature data to deduce fragmentation pathways. ResultsA total of 122 compounds were identified in the reference sample of Jinshui Liujunjian， including 47 flavonoids， 5 amino acids， 13 iridoids， 16 triterpenoid saponins， etc.， of which 42 compounds were confirmed by comparison with reference substances. A total of 21 prototype components were identified in blood components； 50 prototype components were identified in different tissues， among which 13， 10， 7， 21， 11， 6， 14， and 40 prototype components were identified in the heart， liver， spleen， lung， kidney， brain， large intestine， and stomach， respectively. Among them， 7 compounds such as ferulic acid， glycyrrhizic acid， and nobiletin were exposed in the target organs of lung and kidney. ConclusionThis study elucidates the material basis of the reference samples of Jinshui Liujunjian， primarily composed of flavonoids and triterpenoid saponins， along with their in vivo distribution characteristics. These findings provide a scientific basis for establishing quality evaluation indicators and offer references for subsequent pharmacodynamic and pharmacokinetic investigations.

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 summarize and analyze the main clinical characteristics,feature and composition changes of gut microbiota in elderly patients with severe pneumonia,and to further explore the potential correlation between the gut characteristics and the etiology of severe pneumonia in elderly patients.Methods Patients with severe pneu-monia admitted to the respiratory intensive care unit of a tertiary teaching hospital in Changsha were selected as the research subjects.Patients aged ≥65 years were assigned to the elderly severe pneumonia group,while those aged＜65 years were assigned to the non-elderly severe pneumonia group.Based on clinical characteristics and pathogen detection of lower respiratory secretion,the elderly severe pneumonia group was further divided into a pulmonary bacterial infection group and a pulmonary fungal infection group.The pulmonary bacterial infection group was sub-divided into Gram-positive bacteria group and Gram-negative bacteria group based on Gram-staining results.Clinical data of patients were collected,and fecal specimens within 24 hours after admission were obtained for 16S rRNA se-quencing.Differences in gut microbiota characteristics between two groups of patients were compared,and the cor-relation between clinical characteristics of patients in the elderly severe pneumonia group and the abundance of dif-ferential microbiota was analyzed.Subsequently,the gut microbiota characteristics of elderly patients in severe pneumonia group infected by different pathogens were analyzed.Results Gut microbiota analysis showed no signifi-cant statistical differences in α-and β-diversity indicices between patients in the elderly and non-elderly severe pneu-monia groups(both P＞0.05).Linear discriminant analysis effect size(LEFSe)analysis indicated that,compared with patients in the non-elderly severe pneumonia group,the relative abundance of opportunistic pathogens,inclu-ding Pseudomonadales,Moraxellaceae,and Acinetobacter,was significantly higher in patients in the elderly severe pneumonia group(all P＜0.05).Some differential gut microbiota in two groups of patients were correlated with clinical indicators in patients in the elderly severe pneumonia group(all P＜0.05).β-diversity analysis(principal co-ordinate analysis)combined with Anosim analysis revealed that in patients in elderly severe pneumonia group,there was significant differences in gut colony structures between patients in the bacterial and fungal infection groups(R=0.149,P=0.02).Compared with the fungal infection group,patients in bacterial infection group showed a signifi-cantly reduced abundance of probiotics,including Verrucomicrobiales and Collinsella,and opportunistic pathogens such as Akkermansia(all P＜0.05).Conclusion Elderly patients with severe pneumonia have a dysregulated gut microbiota with significantly increased abundance of pathogenic bacteria compared with non-elderly patients.Differ-ential gut microbiota of two groups of patients are correlated with some infection-related and organ function indica-tors in elderly patients with severe pneumonia.Compared with elderly patients with severe fungal pneumonia,those with severe bacterial pneumonia have significant differences in gut colony structures and a notably reduction in probi-otics abundance.

Non-suicidal self-injury(NSSI)refers to a mental disorder characterized by deliberate self-harm behaviors without suicidal intent.The pathogenesis of NSSI remains unclear,but may be related to abnormalities in reward circuitry,pain processing,emotion regulation,impulse control and hypothalamic-pituitary-adrenal axis.Task state functional MRI(fMRI)provides valuable insights into the intrinsic connections and neural circuits mechanisms underlying NSSI during various task states or behaviors.Research progresses of task state fMRI in pathogenesis of NSSI were reviewed in this article.

BACKGROUND:Cervical instability is a common orthopedic disease in young and middle-aged people,and is the early manifestation of cervical spondylosis,which has a great impact on the quality of life of patients.Therefore,early diagnosis of cervical instability to implement early intervention has positive clinical and social significance.OBJECTIVE:The clinical prediction model of cervical instability in young and middle-aged people was constructed based on machine learning to realize early screening of cervical instability in young and middle-aged people before X-ray examination.METHODS:From September 2022 to October 2023,155 young and middle-aged adults with cervical instability and 88 with non-cervical instability recruited through recruitment advertisements and spinal department outpatient of Wangjing Hospital,China Academy of Chinese Medical Sciences were selected as research subjects.The research subjects'general information,living and working habits,discomfort symptoms,visual analog scale score,Neck Disability Index,and 36-ltem Short Form Health Survey were collected on site based on questionnaires.The above information was used as predictive factors.After screening,six machine learning algorithms of Support Vector Machine,LightGBM,RandomForest,Logistic,AdaBoost,and XGBClassifier were used to train the model by ten-fold cross-validation method,and the clinical prediction model of cervical instability was constructed.Area under the curve was used as the main evaluation index.Univariate analysis was performed on the predictors,and SHAP method was used to rank the importance of the predictors.Correlation heat maps were used to show the degree of linear correlation between the predictors and the cervical instability.RESULTS AND CONCLUSION:(1)Among the six machine learning models,RandomForest model was chosen as the final prediction model,including nine predictors,such as age,body mass index,neck circumference/neck length,visual analog scale score,Neck Disability Index,bodily pain,general health,vitality,and mental health,area under the curve=0.725 4,and the calibration degree was good.It could be used as a reference tool for early screening of cervical instability in young and middle-aged people.(2)There were significant differences in age,visual analog scale score,Neck Disability Index,bodily pain,general health,and vitality between the two groups(P＜0.05).(3)The order of importance of predictors was age,Neck Disability Index,visual analog scale score,general health,body mass index,vitality,bodily pain,neck circumference/neck length,mental health,among which age,visual analog scale score,Neck Disability Index were positively correlated with cervical instability,while general health,body mass index,vitality,bodily pain,neck circumference/neck length,and mental health were negatively correlated with cervical instability.

Objective:To explore the application effect of BOPPPS teaching model in the teaching of Food Safety Supervision and Management course.Methods:Students of two parallel classes majoring in the Food Quality and Safety of the same grade of a university in Liaoning Province were selected,and the traditional teaching model and BOPPPS teaching model were used for teaching practice respectively.The scores,teaching effect and satisfaction of two groups were compared.Results:The regular performance and total scores of the experimental group were significantly higher than those of the control group(P＜0.05).In terms of enhancing students'interest in food safety supervision,improving knowledge and skills,cultivating self-directed learning and teamwork abilities,strengthening the close connection between the course and real cases,and providing assistance for future career,the experimental group performed significantly better than the control group(P＜0.05).Moreover,the satisfaction of the experimental group was higher than that of the control group(P＜0.05).Conclusion:BOPPPS teaching model has a remarkable effect in the teaching of Food Safety Supervision and Management course,providing an effective approach for the teaching reform of this course.

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 physicochemical and immune efficacy of filamentous hemagglutinin(FHA)and pertactin(PRN)before and after detoxification treatment in acellular pertussis vaccine.Methods:Thermal stability of FHA and PRN was analyzed by differential scanning fluorescence(DSF)to evaluate melting temperature(Tm)changes of antigen before and after detoxi-fication treatment.Immune efficacy of vaccine containing single FHA or PRN component and acellular pertussis vaccine with pertussis toxin(PT)was assessed by modified intracerebral challenge assay(MICA)and pertussis serological potency test(PSPT).ELISpot and flow cytometry were utilized to evaluate cytokines secretion and immune cells level in spleens obtained from immunized mice.Results:Under detoxification treatment Tm of FHA changed from a single value 57℃to both 58℃and 81℃,but Tm of PRN were both 84℃.Detoxification treatment had no obvious effects on immune efficacy and IL-4/IFN-γ secretion in spleens from mice.In addi-tion,there was no significant difference in splenic immune cells such as CD4+T,CD8+T and CD44+CD62L+memory cells.Conclu-sion:Detoxification treatment of FHA and PRN have no effect on immune efficacy of acellular pertussis vaccine,except for impact on Tm of FHA.

Programmed cell death is an important defense mechanism of the body, which monitors internal lesions and defends external infections to maintain the stability of the body.In the past, there were two main types of cell death: apoptosis and necrosis.With the development of scientific research, new cell death modes, such as pyroptosis and ferroptosis, have attracted much attention and are involved in the occurrence and development of diabetic retinopathy (DR).Many pathogenic factors, such as oxidative stress, hyperglycemia and inflammation, lead to the death of retinal cells in different ways, such as apoptosis, pyroptosis and ferroptosis.Apoptosis is the earliest pathological response of DR, which occurs at the initial stage of oxidative stress, and the integrity of cell membranes is maintained at the early stage of apoptosis.Apoptotic cells are cleared by phagocytes in the tissue before dissolution, which avoids unnecessary inflammation.Pyroptosis is activated by Caspase-1, then cleaves GSDMD, which can lead to pore formation and osmotic cell lysis, resulting in inflammation and immune response.Ferroptosis involves lipid peroxidation and abnormal iron homeostasis.Lipid peroxidation and glutathione depletion are the main markers of ferroptosis.The three cell death modes are independent and interrelated, and play a role in the occurrence and development of diseases together with other cell death modes.This article reviews the current status of cell apoptosis, pyroptosis and ferroptosis in DR.