Objective To explore the mechanism of Tongdu Xingshen acupuncture,the clinical efficacy,systemic inflammatory response,blood-brain barrier and intestinal flora in patients with cognitive impairment after ischemic stroke(IS)were studied.Methods Thirty patients(3 cases shedding)with cognitive impairment after IS were included as the disease group,including patients before treatment as the disease group,patients after Tongdu Xingshen acupuncture treatment as the electroacupuncture group,and 30 healthy controls(3 cases shedding)were included as the healthy group.In the electroacupuncture group,on the basis of the basic treatment,Tongdu Xingshen acupuncture was applied,which was 30 min each time,once a day for 14 days.The MMSE,MoCA and MBI scores of the three groups were observed.The fecal and serum samples from all study subjects were collected,and 16S rDNA sequencing technology and ELISA were used to detect the changes of proinflammatory factors IL-6,IL-1β,TNF-α and S100β in serum in intestinal flora and feces.Results Compared with the healthy group,the MMSE,MoCA,and MBI score of patients in the disease group decreased significantly(P<0.05),serum proinflammatory factors and S100β protein content increased significantly(P<0.05),and the Shannon index(P<0.01)and Simpson index(P<0.001)increased significantly.Compared with the disease group,the MMSE,MoCA,and MBI score of the EA group increased significantly(P<0.05),the serum levels of proinflammatory factors and S100β decreased significantly(P<0.05),Shannon index and Simpson index decreased(P>0.05).The dominant bacterial flora in the healthy group mainly included Bacteroides,Bifidobacterium,Bacteroides,Faecalibacterium,Bifidobacteriaceae,Ruminococcaceae,and Bacteroides and other beneficial bacteria(P<0.05).The dominant flora in the disease group included Proteobacteria,Enterobacteriaceae,Escherichia,Klebsiella and other opportunistic bacteria(P<0.05),while the dominant flora in the EA group was consistent with the healthy group,the relative abundance of beneficial bacteria increased significantly(P<0.05),and the relative abundance of opportunistic bacteria decreased significantly(P<0.05).Spearman correlation analysis found that beneficial bacteria were positively correlated with clinical efficacy related indicators,but with serum proinflammatory factors and the content of S100β was negatively correlated.Conclusion Tongdu Xingshen acupuncture can regulate the diversity of intestinal flora to increase the abundance of Bacteroides,Bifidobacterium,Faecalibacterium,and other beneficial bacteria,regulate the intestinal microecological balance,Thereby regulating systemic inflammation and blood-brain barrier function,which plays a role in improving cognitive function.

OBJECTIVE: To observe the effects of electroacupuncture (EA) at "Shenting" (GV24) and "Baihui" (GV20) on mitochondrial autophagy in hippocampal neurons and silent information regulator sirtuin 1 (Sirt1)/forkhead box O3 (FOXO3)/PTEN-inducible kinase 1 (PINK1)/Parkin pathway in rats with learning-memory impairment after cerebral ischemia reperfusion injury. METHODS: A total of 35 male SD rats were randomly divided into a sham operation group (9 rats) and a modeling group (26 rats). In the modeling group, middle cerebral artery occlusion method was used to establish the middle cerebral artery ischemia-reperfusion (MCAO/R) model, and 18 rats of successful modeling were randomly divided into a model group and an EA group, 9 rats in each one. EA was applied at "Shenting" (GV24) and "Baihui" (GV20) in the EA group, 30 min a time, once a day for 14 days. After modeling and on 7th and 14th days of intervention, neurologic deficit score was observed; the learning-memory ability was detected by Morris water maze test; the morphology of neurons in CA1 area of hippocampus was detected by Nissl staining; the mitochondrial morphology was observed by transmission electron microscopy; the protein expression of Beclin-1, microtubule-associated protein 1 light chain 3B (LC3B), P62, Sitrt1, FOXO3, PINK1 and Parkin was detected by Western blot. RESULTS: After modeling, the neurologic deficit scores in the model group and the EA group were higher than that in the sham operation group (P<0.001); on 7th and 14th days of intervention, the neurologic deficit scores in the model group were higher than those in the sham operation group (P<0.001), the neurologic deficit scores in the EA group were lower than those in the model group (P<0.05, P<0.01). After modeling, the escape latency in the model group and the EA group was prolonged compared with that in the sham operation group (P<0.001); on 9th-13th days of intervention, the escape latency in the model group was prolonged compared with that in the sham operation group (P<0.001), the escape latency in the EA group was shortened compared with that in the model group (P<0.05, P<0.01, P<0.001). The number of crossing plateau in the model group was less than that in the sham operation group (P<0.001); the number of crossing plateau in the EA group was more than that in the model group (P<0.05). In the model group, in CA1 area of hippocampus, the number of neurons was less, with sparse arrangement, nuclear fixation, deep cytoplasmic staining, and reduction of Nissl substance; the morphology of mitochondrion was swollen, membrane structure was fragmented, and autophagic lysosomes were formed. Compared with the model group, in the EA group, in CA1 area of hippocampus, the number of neurons was increased, the number of cells of abnormal morphology was decreased, and the number of Nissl substance was increased; the morphology of mitochondrion was more intact and the number of autophagic lysosomes was increased. Compared with the sham operation group, in the model group, the protein expression of Beclin-1, FOXO3, PINK1, Parkin and the LC3BⅡ/Ⅰ ratio in hippocampus were increased (P<0.01, P<0.001), while the protein expression of P62 was decreased (P<0.05). Compared with the model group, in the EA group, the protein expression of Beclin-1, Sirt1, FOXO3, PINK1, Parkin and the LC3BⅡ/Ⅰratio in hippocampus were increased (P<0.001, P<0.01), while the protein expression of P62 was decreased (P<0.001). CONCLUSION EA at "Shenting" (GV24) and "Baihui" (GV20) can relieve the symptoms of neurological deficits and improve the learning-memory ability in MCAO/R rats, its mechanism may relate to the modulation of Sirt1/FOXO3/PINK1/Parkin pathway and the enhancement of mitochondrial autophagy.
Animals ; Electroacupuncture ; Male ; Rats, Sprague-Dawley ; Rats ; Forkhead Box Protein O3/genetics* ; Reperfusion Injury/metabolism* ; Ubiquitin-Protein Ligases/genetics* ; Brain Ischemia/complications* ; Mitochondria/genetics* ; Autophagy ; Protein Kinases/genetics* ; Sirtuin 1/genetics* ; Humans ; Memory Disorders/psychology* ; Signal Transduction

The specific mechanisms of interferon regulatory factor 8(IRF8)in porcine intestinal in-nate immunity and resistance to enteric virus infection remain to be elucidated.To investigate the immunoregulatory role of IRF8,establishing an IRF8 gene knockout porcine intestinal epithelial cell(IPEC-J2)monoclonal cell line is of significant importance.This study initially aimed to obtain recombinant adeno-associated virus rAAV-sgIRF8-eGFP capable of knocking out the IRF8 gene through co-transfection of HEK-293T cells with three plasmids.Subsequently,IPEC-J2 cells were infected with the virus,and those expressing eGFP were selected by flow cytometry and cultured to form monoclonal cell lines.These cell lines were then identified by Sanger sequencing and West-ern blot techniques.Lastly,qPCR analysis was used to measure the expression levels of interferon factors IFN-α,IFN-β,IFN-γ and IFN-λ,providing preliminary insights into the impact of IRF8 gene knockout on IPEC-J2 cell immunity.The results demonstrated successful generation of rAAV-sgIRF8-eGFP,which successfully infected IPEC-J2 cells leading to eGFP fluorescence.Flow cytometry followed by cell culture led to the establishment of two monoclonal cell lines,IRF8-KO1 and IRF8-KO3.Sanger sequencing revealed a five-base deletion in IRF8-KO1 and a seven-base dele-tion in IRF8-KO3.Western blot confirmed the absence of IRF8 protein expression in IRF8-KO1,making it an ideal candidate monoclonal cell line.qPCR analysis of interferon factors indicated sig-nificant decrease in IFN-γ(P＜0.05)and IFN-λ(P＜0.01)transcription level in IRF8-knockout cells,while the transcription levels of IFN-α and IFN-β remained relatively unchanged.This study successfully established an IRF8 gene knockout IPEC-J2 monoclonal cell line,providing a founda-tion for further research on IRF8-related porcine intestinal immune regulation and mechanisms of intestinal virus infection.

The specific mechanisms of interferon regulatory factor 8(IRF8)in porcine intestinal in-nate immunity and resistance to enteric virus infection remain to be elucidated.To investigate the immunoregulatory role of IRF8,establishing an IRF8 gene knockout porcine intestinal epithelial cell(IPEC-J2)monoclonal cell line is of significant importance.This study initially aimed to obtain recombinant adeno-associated virus rAAV-sgIRF8-eGFP capable of knocking out the IRF8 gene through co-transfection of HEK-293T cells with three plasmids.Subsequently,IPEC-J2 cells were infected with the virus,and those expressing eGFP were selected by flow cytometry and cultured to form monoclonal cell lines.These cell lines were then identified by Sanger sequencing and West-ern blot techniques.Lastly,qPCR analysis was used to measure the expression levels of interferon factors IFN-α,IFN-β,IFN-γ and IFN-λ,providing preliminary insights into the impact of IRF8 gene knockout on IPEC-J2 cell immunity.The results demonstrated successful generation of rAAV-sgIRF8-eGFP,which successfully infected IPEC-J2 cells leading to eGFP fluorescence.Flow cytometry followed by cell culture led to the establishment of two monoclonal cell lines,IRF8-KO1 and IRF8-KO3.Sanger sequencing revealed a five-base deletion in IRF8-KO1 and a seven-base dele-tion in IRF8-KO3.Western blot confirmed the absence of IRF8 protein expression in IRF8-KO1,making it an ideal candidate monoclonal cell line.qPCR analysis of interferon factors indicated sig-nificant decrease in IFN-γ(P＜0.05)and IFN-λ(P＜0.01)transcription level in IRF8-knockout cells,while the transcription levels of IFN-α and IFN-β remained relatively unchanged.This study successfully established an IRF8 gene knockout IPEC-J2 monoclonal cell line,providing a founda-tion for further research on IRF8-related porcine intestinal immune regulation and mechanisms of intestinal virus infection.

Objective To explore the mechanism of Tongdu Xingshen acupuncture,the clinical efficacy,systemic inflammatory response,blood-brain barrier and intestinal flora in patients with cognitive impairment after ischemic stroke(IS)were studied.Methods Thirty patients(3 cases shedding)with cognitive impairment after IS were included as the disease group,including patients before treatment as the disease group,patients after Tongdu Xingshen acupuncture treatment as the electroacupuncture group,and 30 healthy controls(3 cases shedding)were included as the healthy group.In the electroacupuncture group,on the basis of the basic treatment,Tongdu Xingshen acupuncture was applied,which was 30 min each time,once a day for 14 days.The MMSE,MoCA and MBI scores of the three groups were observed.The fecal and serum samples from all study subjects were collected,and 16S rDNA sequencing technology and ELISA were used to detect the changes of proinflammatory factors IL-6,IL-1β,TNF-α and S100β in serum in intestinal flora and feces.Results Compared with the healthy group,the MMSE,MoCA,and MBI score of patients in the disease group decreased significantly(P<0.05),serum proinflammatory factors and S100β protein content increased significantly(P<0.05),and the Shannon index(P<0.01)and Simpson index(P<0.001)increased significantly.Compared with the disease group,the MMSE,MoCA,and MBI score of the EA group increased significantly(P<0.05),the serum levels of proinflammatory factors and S100β decreased significantly(P<0.05),Shannon index and Simpson index decreased(P>0.05).The dominant bacterial flora in the healthy group mainly included Bacteroides,Bifidobacterium,Bacteroides,Faecalibacterium,Bifidobacteriaceae,Ruminococcaceae,and Bacteroides and other beneficial bacteria(P<0.05).The dominant flora in the disease group included Proteobacteria,Enterobacteriaceae,Escherichia,Klebsiella and other opportunistic bacteria(P<0.05),while the dominant flora in the EA group was consistent with the healthy group,the relative abundance of beneficial bacteria increased significantly(P<0.05),and the relative abundance of opportunistic bacteria decreased significantly(P<0.05).Spearman correlation analysis found that beneficial bacteria were positively correlated with clinical efficacy related indicators,but with serum proinflammatory factors and the content of S100β was negatively correlated.Conclusion Tongdu Xingshen acupuncture can regulate the diversity of intestinal flora to increase the abundance of Bacteroides,Bifidobacterium,Faecalibacterium,and other beneficial bacteria,regulate the intestinal microecological balance,Thereby regulating systemic inflammation and blood-brain barrier function,which plays a role in improving cognitive function.

Objective:To analyze the relevant records of Taibai (SP3) in ancient literature based on data mining technology; To summarize its application law.Methods:Ancient book materials included in the Zhong Hua Yi Dian (5th edition) were retrieved. Relevant information of Taibai (SP3) such as main treatment syndrome and compatibility of acupoints was extracted, and a database was established. The frequency of the data was counted. Gephi 0.10.1 software and SPSS Statistics 21.0 statistical software were used to carry out visualization analysis and clustering analysis on data. Results:A total of 248 articles were included, involving 43 ancient books, including 65 prescriptions for single acupoint application and 183 prescriptions for compatibility application. The main treatment of Taibai (SP3) involved internal medicine, surgery, gynecology and infectious diseases. There were 35 kinds of main treatment of single acupoint, and 12 kinds of dominant diseases, including vomiting, fever and dysentery. There were 47 kinds of main diseases and 14 kinds of dominant diseases such as fever, heartache and constipation. There were 138 acupoints, most of which were specific acupoints, mainly the spleen meridian, and the high frequency acupoints were Zusanli (ST36), Dadu (SP2), Gongsun (SP4) and so on. Clustering analysis obtained 8 kinds of acupoint combinations, which were potential prescriptions for Taibai (SP3).Conclusion:Taibai (SP3) was widely used in clinical practice in ancient times, with 8 kinds of diseases such as fever, constipation and abdominal pain as the main dominant diseases, and most of them were compatible with specific acupoints such as Zusanli (ST36), Dadu (SP2), Gongsun (SP4).

The efficacy and safety of venetoclax combined with reduced dose HAD regimen in the treatment of newly diagnosed acute myeloid leukemia (AML) was investigated. From May 2022 to January 2023, a total of 25 patients with newly diagnosed AML were treated with venetoclax combined with reduced-dose HAD regimen as induction therapy. Accoding to the 2017 ELN recommendations, 13 (52.0%) in favoable, 3 (12.0%) in intemediate, and 9 (36.0%) in adverse. The ORR (CR rate+PR rate) was 88.0%, and the CR rate was 84.0%. By May 30, 2023, with a median follow-up of 9 months, 1 year overall survival, event-free survival, and relapse-free survival were 100%, 94.7%, and 94.7%, respectively. All patients received 1-5 cycles of consolidation therapy and two median cycles. Treatment with venetoclax and reduced dose of HAD regimen in the treatment of patients with newly diagnosed AML was high effective and safe.

Objective:To analyze the ultrasonographic measurements of inferior vena cava (IVC) and abdominal aorta in healthy full-term neonates throughout the early postnatal period.Methods:Prospective observational study was conducted. A total of 132 healthy full-term neonates, who were born at the Kunshan First People′s Hospital between May 1 st and August 30 th, 2023, were enrolled as the study subjects. Two-dimensional and M-mode ultrasonography were used to measure the maximum and minimum diameters of the IVC and abdominal aorta in the early postnatal period. The IVC collapsibility index, the ratio of maximum IVC diameter to abdominal aorta diameter, and the ratio of minimum IVC diameter to abdominal aorta diameter were calculated. These neonates were stratified by gender, birth mode, gestational age and birth weight (<3 000 or ≥3 000 g), and independent sample t-test or Kruskal-Wallis H test was used to compare the ultrasonography measurements by different groups. Results:Among the 132 neonates, 81 were males, with a gestational age of (39.2±1.0) weeks, and a birth weight of (3 326±409) g. There were no significant statistical differences in the the maximum and minimum diameters of the IVC and abdominal aorta assessed by both two-dimensional and M-modes between the first and second days (all P>0.05). Additionally, no statistical differences were observed in the ultrasonographic measurements among neonates of different sexes, birth modes, and gestational ages (all P>0.05); but there were statistically differences between the group with birth weight of <3 000 g and the group with birth weight of ≥3 000 g (all P<0.05). There were no statistically differences in IVC collapsibility index, the ratio of the maximum diameter of IVC to the diameter of abdominal aorta, and the ratio of the minimum diameter of IVC to the diameter of abdominal aorta between the group with birth weight of <3 000 g and the group with birth weight of≥3 000 g (all P>0.05). Conclusions:The diameters of the IVC and abdominal aorta in healthy full-term neonates during the early postnatal period are correlated with birth weight. The IVC collapsibility index and the ratio of IVC diameter to abdominal aorta diameter are unrelated to birth weight and can be used to assess newborn blood volume or right cardiac preload.

Objective:To discuss dominant symptoms and compatibility rules of Dazhui(GV14) based on data mining.Methods:Literature related to Dazhui (GV14) was retrieved from CNKI, Wangfang, VIP, China Biomedical Literature Database (CBM) and Pubmed databases from January 1, 2012 to August 15, 2022, and the main symptoms of Dazhi (GV14) and the compatibility of acupoints were summarized. Gephi 0.9.5 software was used for complex network analysis to compare the treatment for dominant symptoms with single acupoint of Dazhi (GV14) and the compatibility of the acupoint. SPSS Modeler 18.0 software was used to analyze the association rules of acupoint combination based on Apriori algorithm. The clustering analysis of high frequency acupoints was carried out by SPSS Statistics 26.0 software.Results:A total of 722 articles were included, involving 732 prescriptions. The dominant symptoms of single acupoint were cervical spondylosis, acne, and cold; the treatment for dominant symptoms with compatibility included 14 types, such as cervical spondylosis, allergic rhinitis, ischemic stroke sequelae. The meridian compatibility was dominated by bladder meridian, and the frequency of yang meridians was higher than yin meridians. The compatibility of specific acupoints such as Xiahe acupoint, Beishu acupoint and Bahui acupoint were the main acupoints, and the high frequency acupoints were 33 acupoints such as Feishu (BL13), Baihui (GV20), Fengchi (GB20) and Zusanli (ST36), obtaining 4 series and 8 types of compatible combinations of Dazhui (GV14).Conclusions:Dazhui (GV14) is widely used in the treatment of internal diseases, such as respiratory diseases, nervous system diseases and vertebral artery type of cervical spondylosis. It tends to be flexibly used with multiple compatibility and clustering combination of specific acupoints.

Objective: To prepare a composite membrane by chitosan/β-sodium glycerophosphate(CS/β-GP) thermosensitive hydrogel combined with stromal cell derived factor-1(SDF-1) and observe its biological characteristics in vitro. Methods: Different doses of SDF-1 were added into CS/β-GP solution and then the thermosensitive gel time was measured. The SDF-1/CS/β-GP solution was membrane paved and dried to prepare composite membranes. The morphological characteristics were observed by scanning electron microscope(SEM). Composite membranes were placed into cell culture medium, and the supernatant(n=3) was extracted after standing at 6, 12, 24, 36, 48, 60 h, respectively. The concentration of SDF-1 in the solution was measured. Bone mesenchymal stem cells(BMSCs) were cultured in the Transwell room, and the composite membranes containing different concentrations of SDF-1 were placed in the lower chamber. There were four groups(n=3): Group M0 used CS/β-GP membrane(control group), Group M1, M2, M3 used SDF-1/CS/β-GP membrane(SDF-1 was 100, 200, 400 ng/mL respectively). After culture for 6, 12 and 24 h, the cells under the membrane were preserved and Giemsa stained and counted. The absorbance(OD) value was measured by MTT method to calculate the cell proliferation rate. SPSS 19.0 was used for multi-factor analysis of variance. Results : After adding a certain amount of SDF-1 into CS/β-GP solution, the gel time did not change significantly(P>0.05). The SDF-1/CS/β-GP membrane was translucent and porous at 37 ℃. In this experiment, the volumic mass of SDF-1 released by SDF-1/CS/β-GP composite membrane increased gradually with the experimental time(P<0.01). Transwell cell chemotaxis test showed that the number of BMSCs cells with directional migration increased with the prolongation of observation time(P<0.01) and the increase of SDF-1 volumic mass(P<0.01). In MTT test, the OD value of migration cell solution increased with the prolongation of time(P<0.01) and the increase of SDF-1 volumic mass(P<0.01). Conclusion The SDF-1/CS/β-GP composite membrane has a porous structure and biological activity of chemotactic BMSCs directional migration. It is a potential membrane for guided tissue regeneration.