Objective To establish a rat model of Alzheimer's disease (AD) expressing human triple mutant amyloid precursor protein (APP) in the hippocampus, and to provide a model for the study of disease mechanisms and drug development. Methods Twenty-four 12-week-old SPF-grade female SD rats were randomly divided into a blank control group, a virus control group and an experimental group, with eight rats in each group; among them, the experimental group received a stereotaxic injection of adeno-associated virus (AAV) carrying the human triple mutant APP and NanoLuc luciferase genes into the hippocampus. In vivo imaging was used to observe viral expression in the brains of rats in each group, the novel object recognition test was used to assess the recognition memory of the rats in each group, real-time fluorescent quantitative PCR was used to detect the expression level of the APP gene, HE staining was used to examine the brain histopathology, Nissl staining was used to assess the hippocampal lesions, and immunohistochemistry was used to detect the deposition of amyloid β-protein (Aβ). Results In vivo imaging showed that reporter fluorescence was detected in the brains of rats in both experimental and virus control groups. Fluorescence quantitative PCR showed that the expression level of the APP gene was significantly increased in the brains of rats in the experimental group (P<0.01). Novel object recognition test revealed that the recognition memory of rats in the experimental group was significantly reduced compared with that of the blank control group (P<0.01). Six months after recombinant AAV virus infection, HE staining and Nissl staining of brain tissues showed that the number of neurons and Nissl bodies in the CA1 region of the hippocampus in the experimental group was reduced and disorganized; immuno-histochemistry testing of the CA1 region of the hippocampus and the pyramidal cell layer of the experimental group revealed prominent brown deposits, indicating Aβ protein deposition. Conclusion The rat model successfully established by stereotaxic injection and AAV-mediated delivery of human triple mutant APP gene exhibits typical AD features, providing a valuable animal model for studying AD pathology and developing drug therapies targeting Aβ protein deposition.

Objective To establish a rat model of Alzheimer's disease (AD) expressing human triple mutant amyloid precursor protein (APP) in the hippocampus, and to provide a model for the study of disease mechanisms and drug development. Methods Twenty-four 12-week-old SPF-grade female SD rats were randomly divided into a blank control group, a virus control group and an experimental group, with eight rats in each group; among them, the experimental group received a stereotaxic injection of adeno-associated virus (AAV) carrying the human triple mutant APP and NanoLuc luciferase genes into the hippocampus. In vivo imaging was used to observe viral expression in the brains of rats in each group, the novel object recognition test was used to assess the recognition memory of the rats in each group, real-time fluorescent quantitative PCR was used to detect the expression level of the APP gene, HE staining was used to examine the brain histopathology, Nissl staining was used to assess the hippocampal lesions, and immunohistochemistry was used to detect the deposition of amyloid β-protein (Aβ). Results In vivo imaging showed that reporter fluorescence was detected in the brains of rats in both experimental and virus control groups. Fluorescence quantitative PCR showed that the expression level of the APP gene was significantly increased in the brains of rats in the experimental group (P<0.01). Novel object recognition test revealed that the recognition memory of rats in the experimental group was significantly reduced compared with that of the blank control group (P<0.01). Six months after recombinant AAV virus infection, HE staining and Nissl staining of brain tissues showed that the number of neurons and Nissl bodies in the CA1 region of the hippocampus in the experimental group was reduced and disorganized; immuno-histochemistry testing of the CA1 region of the hippocampus and the pyramidal cell layer of the experimental group revealed prominent brown deposits, indicating Aβ protein deposition. Conclusion The rat model successfully established by stereotaxic injection and AAV-mediated delivery of human triple mutant APP gene exhibits typical AD features, providing a valuable animal model for studying AD pathology and developing drug therapies targeting Aβ protein deposition.

Objective To investigate the role and related mechanisms of exosomes derived from mesenchymal stem cells (MSCs) in radiation-induced lung injury (RILI). Methods Human umbilical cord-derived MSCs were isolated and cultured for the extraction and identification of exosomes. Eighteen male SD rats were randomly divided into Control group, RILI group and RILI + exosomes group (EXO group), with 6 rats in each group. Except for Control group, the other groups received a single X-ray dose of 30 Gy to the right lung. Immediately after irradiation, the EXO group was administered 2 × 10⁹ exosomes/kg via tail vein injection. Control group and RILI group were given the same volume of normal saline. Eight weeks post-irradiation, the rats were sacrificed, lung tissue and peripheral venous blood were collected. HE and Masson staining were employed to observe the pathological and fibrotic changes of lung tissue. The levels of serum inflammatory factors IL-6, IFN-γ, TNF-α, and IL-10 were detected by ELISA. RT-qPCR was used to assess the mRNA levels of IL-1β, IL-6, Cdh1, and Col1a1 in lung tissue. The expression levels of Vimentin and TGF-β1 in lung tissue were measured by immunohistochemical staining. The expression levels of AMPK, p-AMPK, and TGF-β1 in lung tissue were detected by Western blot. Results MSC-derived exosomes were successfully extracted and identified. Compared with RILI group, EXO group showed significantly reduced pathological changes of lung inflammation and collagen deposition. The levels of serum inflammatory factors IL-6, INF-γ, and TNF-α were significantly decreased (P < 0.05), and the level of anti-inflammatory factor IL-10 was significantly increased (P < 0.05). The mRNA levels of IL-1β, IL-6, and Col1a1 in lung tissue were significantly decreased (P < 0.05 or P < 0.01), and the mRNA level of Cdh1 was significantly increased (P < 0.05 or P < 0.01). The levels of Vimentin and TGF-β1 in lung tissue were significantly reduced, while p-AMPK level was significantly up-regulated (P < 0.05). Conclusion Exosomes derived from MSCs may alleviate RILI by inhibiting inflammatory responses and regulating epithelial-mesenchymal transition mediated by AMPK/TGF-β1 signaling pathway.

Polysaccharides are the important material basis of traditional Chinese medicine（TCM）， and have various pharmacological activities such as immunomodulation， antitumor and anti-aging. Due to the large molecular weight of TCM polysaccharides， their structural analysis and oral absorption mechanism are facing technical challenges， and the current research on their structure-activity relationships has made some breakthroughs， while the research on their oral absorption mechanisms is relatively slow. In-depth study of the oral absorption mechanism of TCM polysaccharides is not only crucial for the interpretation of their action pathways and efficacy in vivo， but also helpful for the interpretation of their pharmacological effects， rational clinical applications and the discovery of new targets. In recent years， the application of fluorescent labeling and isotopic labeling methods has provided new technical means for the oral absorption studies of polysaccharides， which has promoted the development of oral absorption studies of TCM polysaccharides. In this paper， we reviewed the oral absorption pathways and labeling techniques of TCM polysaccharides， and concluded that they can be absorbed orally through transmembrane， cellular bypass， and M-cell-mediated transport， of which transmembrane pathway is the main absorption pathway， and summarized the labeling reactions of four fluorescent labeling and isotopic labeling methods with TCM polysaccharides， which can provide references for evaluating the absorption pathways of TCM polysaccharides， screening active TCM polysaccharides， establishing pharmacodynamic models and comprehensively elucidating the mechanism of TCM polysaccharides.

Objective To observe the clinical efficacy of acupuncture combined with rehabilitation training in treating qi deficiency and blood stasis type of hypertensive cerebral hemorrhage in the recovery stage.Methods A total of 132 patients with qi deficiency and blood stasis type of hypertensive cerebral hemorrhage in the recovery period were randomly divided into observation group and control group,with 66 cases in each group,the control group was given western medicine conventional treatment combined with rehabilitation training,and the observation group was treated with acupuncture on the basis of the control group.Both groups of patients were treated for 12 consecutive weeks.After 12 weeks of treatment,the clinical efficacy of the two groups was evaluated.The changes of simplified Fugl-Meyer Assessment(FMA),National Institutes of Health Neurological Impairment Scale(NIHSS),and traditional Chinese medicine(TCM)syndrome scores,as well as the changes of serum interleukin 6(IL-6),homocysteine(Hcy),and endothelin 1(ET-1),serum matrix metalloproteinase 9(MMP-9),and brain-derived neurotrophic factor(BDNF)levels were observed before and after the treatment of the patients in the two groups.The changes of serum serine-threonine protein kinase(AKT),phosphatidylinositol-3 kinase(PI3K),and Bcl-2-related X protein(bax)levels were compared between the two groups before and after treatment.Results(1)After treatment,the serum IL-6,Hcy,ET-1 levels of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving the serum IL-6,Hcy,ET-1 levels,and the difference was statistically significant(P＜0.05).(2)After treatment,the serum MMP-9 and BDNF levels of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving serum MMP-9 and BDNF levels,with statistically significant differences(P＜0.05).(3)After treatment,the serum AKT,PI3K,bax levels of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving serum AKT,PI3K,bax levels,and the difference was statistically significant(P＜0.05).(4)After treatment,the FMA score,TCM syndrome scores,and NIHSS score of patients in the two groups were significantly improved(P＜0.05),and the observation group was significantly superior to the control group in improving the FMA score,TCM syndrome scores,and NIHSS score,and the differences were statistically significant(P＜0.05).(5)The total effective rate was 93.34%(62/66)in the observation group and 81.82%(54/66)in the control group.The efficacy of the observation group was superior to that of the control group,and the difference was statistically significant(P＜0.05).Conclusion Acupuncture combined with rehabilitation training for the treatment of patients recovering from hypertensive cerebral hemorrhage of qi deficiency and blood stasis type can significantly reduce the patient's inflammatory response,regulate the level of neurofactors,inhibit neuronal apoptosis,and promote the recovery of neurological function,and the clinical efficacy is remarkable.

Objective To explore the application effect of the Mini-CEX evaluation model based on the OBE concept in the clinical Practice teaching of neurology.Methods We Selected 100 students who will Practice in the Department of Neurology from 2022 to 2023 as the research objects,and divided them into the experimental group(n=50)and the control group(n=50).Under the guidance of the OBE concept,the experimental group was guided by learning outcomes,refined the teaching objectives,and applied the Mini-CEX evaluation mode for evaluation and feedback.In contrast,the control group adopted the traditional teaching mode.Combined with the observation data,we analyzed and compared the data of various indicators of the two groups of students at the beginning and end of the internship.Results At the end of the internship,the scores of clinical consultation,Physical examination,humanistic medicine,clinical diagnosis,health consultation,organizational effect,and overall evaluation of the experimental group were significantly improved and were higher than those of the control group.After the Practice,in terms of skill test scores,the experimental group scored higher than the control group,the difference was statistically significant(P<0.05),and the experimental group also scored higher in satisfaction evaluation than the control group.Conclusion The Mini-CEX evaluation teaching model based on the concept of OBE is applied to the clinical practice teaching of the neurology department,which can enhance the training effect of students'clinical practice skills.

Objective:To investigate the application of Wuxing music combined with traditional Chinese medicine bath and golden paste encapsulation in patients with psoriasis vulgaris(blood-heat syndrome).Methods:In this prospective study,80 patients with psoriasis vulgaris(blood heat syndrome)admitted to our hospital from Nov 2020 to Nov 2022 were included and randomly divided into the control group and the observation group,with 40 cases in each group.The control group received routine nursing intervention,and the observation group received Wuxing music combined with traditional Chinese medicine bath and golden paste encapsulation nursing on the basis of control group,4 weeks as a course of treatment.After 2 courses of nursing,nursing effect,the scores of SCL-90 and DQOLS,the levels of serum inflammatory factors[interleukin-13(IL-13),transforming growth factor-β(TGF-β),and tumor necrosis factor-α(TNF-α)],and nursing satisfaction were compared between the two groups.Results:The area and severity of skin lesions in the two groups were lower than before nursing,and the changes in the observation group were more significant(P＜0.05).The scores of SCL-90 and DQOLS in the two groups were lower than those before nursing,and the changes in the observation group were more significant(P＜0.05).The levels of IL-13,TGF-β and TNF-α in the two groups were lower than those before nursing,and the levels of these indicators in the observation group were significantly lower than those in the control group(P＜0.05).The satisfaction rate of the observation group was higher than that of the control group(P＜0.05).Conclusions:Wuxing music combined with traditional Chinese medicine bath and golden paste encapsulation has a significant nursing effect on patients with psoriasis vulgaris(blood heat syndrome).It can decrease serum inflammatory factors,alleviate the degree of skin lesions,and iprove quality of life and nursing satisfaction.

Objective To comprehensively evaluate the current situation of traditional Chinese medicine resource allocation in grassroots community health service centers in Guangzhou in 2022.Methods Based on the index system of traditional Chinese medicine resource allocation(community health service center)in Guangzhou,MATLAB R2021a and SPSS 27 software were used to comprehensively evaluate the current situation of traditional Chinese medicine resource allocation in 116 community health service centers in Guangzhou by TOPSIS method and RSR method.Results The allocation of TCM resources in 5 communities,including Xiaoguwei Street Community Health Service Center in Panyu District,Guangzhou City,Dadong Street Community Health Service Center in Yuexiu District,Guangzhou City,Fengyuan Street Community Health Service Center in Leiwan District,Guangzhou City,was evaluated as"excellent",and the allocation of TCM resources in 4 communities was rated as"poor".In addition,27,53 and 27 community health service centers were rated as"upper middle","medium"and"lower middle"respectively.Analysis of variance showed that the difference was statistically significant(F = 231.268,P<0.001).Conclusion TOPSIS method combined with RSR method can better evaluate the allocation of TCM resources in grass-roots communities:The allocation of TCM resources in grass-roots communities in Guangzhou is generally good,but there are still significant differences among different communities.In the future,health administrative departments at all levels in Guangzhou can rationally allocate resources according to the differences of different com-munities and better improve the capacity building of traditional Chinese medicine service in grassroots communities.

OBJECTIVE Based on fibroblast activation protein(FAP)gene promoter as the response element,to develop a new dual luciferase reporting system for the screening of drugs related to myocardial fibrosis.METHODS The promoter fragment of mouse FAP gene was synthesized in vitro and cloned into plasmid psiCHECK2 to replace HSV-TK promoter,and then a new recombinant plasmid psiCHECK2-FAP was obtained.After the recombinant plasmid psiCHECK2-FAP was digested by restriction endonucliase Hind Ⅲ,the product digested was identified by agar-gel electrophoresis and sequencing.After psiCHECK2-FAP was transient transfected into mouse cardiac fibroblasts(MCFs),and continued cultured for 24 h,and MCFs were treated with Ransforming growth factor β1(TGF-β1,5 μg·L-1)or angiotensinⅡ(Ang Ⅱ,1 μmol·L-1)or palmitic acid(PA,100 μmol·L-1)for 0,12,24,48 h,respectively,the double luciferase reporter gene assay was used to detect luciferase activity;After psiCHECK2-FAP was transient transfected into MCFs,the cells were pretreated with Dapa(1 μmol·L-1)for 1 h,and supplemented with TGF-β1(5 μg·L-1)or AngⅡ(1 μmol·L-1)or PA(100 μmol·L-1),continued treatment for 24 h,the double luciferase reporter gene assay was used to detect luciferase activity,and the expression of collagenⅠand collagen Ⅲ protein was detected with Western blotting.RESULTS The recombinant plasmid psiCHECK2-FAP was digested into two fragments by Hind Ⅲ with the expected strip size,and the sequencing results were completely consistent with the theoretical sequence;Compared with control group,the collagenⅠand collagen Ⅲ protein expression were significantly increased by TGF-β1 or Ang Ⅱ or PA in MCFs(P<0.05,P<0.01).However,compared with TGF-β1 or Ang Ⅱ or PA group,the intervention of Dapa significantly alleviated the promoter activity of FAP gene and the expression of collagenⅠand collagen Ⅲ protein(P<0.05,P<0.01);Compared with control group,luciferase activity was significantly increased by TGF-β1 or Ang Ⅱ or PA(P<0.05,P<0.01),reaching the peak at 24 h.Compared with TGF-β1 or AngⅡ or PA group,the intervention of Dapa significantly decreased luciferase activity(P<0.05,P<0.01).CONCLUSSION Based on the promoter of FAP gene as the response element,a noval dual luciferase reporter gene system was established and showed a good sensitivity to the promyocardial fibrosis factor in MCFs,which can provide strategies for the development of antimyocar-dial fibrosis drugs.

BACKGROUND:The pedicle navigation template has many advantages,but there are still some problems.For example,poor soft tissue dissection leads to poor adhesion of the pedicle navigation template,resulting in screw path deviation;careful dissection of soft tissue to fit the pedicle navigation template leads to prolonged surgery time and increased bleeding;the design of the pedicle navigation template cannot predict the vertebral rotation and the impact of body position changes,resulting in the poor fitting. OBJECTIVE:To explore the utility of a new 5-point positioning point-contact pedicle navigation template in the case of scoliosis and complex pedicle. METHODS:A total of 20 patients with scoliosis and complicated pedicle admitted to the Department of Spinal Surgery,Guizhou Hospital,Beijing Jishuitan Hospital from February 2020 to February 2023 were selected for scoliosis orthopedics.During the operation,the 5-point positioning point-contact pedicle navigation template was used to guide the screws.According to the inclusion and exclusion criteria,34 cases were matched as the empirical nail placement group,and conventional barehanded nail placement was performed.The time of placement,the amount of bleeding,the number of fluoroscopies,the number of manual diversions,the level and accuracy of pedicle screws,the complications of placement,and the rate of correction of main curvature were compared between the two groups. RESULTS AND CONCLUSION:(1)There were no significant differences in sex,age,coronal Cobb's angle of the main curvature,bending Cobb's angle of the main curvature,pedicle variation,apex rotation,fusion segment,number of screws,level of screws,accuracy of screws,and rate of correction of main curvature between the navigation template group and the empirical nail placement group(P>0.05).(2)Compared with the empirical nail placement group,the navigation template group had more advantages in time of placement(P=0.034),amount of bleeding(P=0.036),number of fluoroscopies(P=0.000)and number of manual diversions(P=0.021).(3)There were 0 cases of screw-related complications in both groups.(4)In conclusion,the 5-point positioning point-contact 3D printing pedicle navigation template has a claw-like structure.It can firmly adapt to various deformities of the lamina articular process,avoid drift,and accurately place the screws.It has a point-like contact lamina structure to avoid extensive and complete dissection of the posterior structure,and reduce bleeding,operation time,and trauma.Pre-designed screw entry points and directions can reduce the number of fluoroscopy and operation time.Segmental design can avoid discomfort due to changes in anesthesia position.The operation is simple and the accuracy of screw placement is high.