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

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 a Chinese pedigree with Hereditary coagulation factor Ⅻ (FⅫ) deficiency duo to variants of F12 gene and explore its molecular pathogenesis. Methods:A patient who underwent laparoscopic cystectomy at the Department of Gynecology of the First Affiliated Hospital of Wenzhou Medical University in June 2012 was selected as the study subject. Coagulation factor indexes of the proband and her family members (5 individuals from three generations) were determined. All exons, flanking sequences, 5′ and 3′ untranslated regions of the F12 gene of the proband and her family members were analyzed by direct sequencing. Three bioinformatics software was used to analyze the conservation, pathogenicity and protein model of the variant. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No. 2012-17). Results:The activated partial thromboplastin time (APTT), FⅫ activity (FⅫ: C) and FⅫ antigen (FⅫ: Ag) of the proband was 180.0 s, 1.0% and 2.1%, respectively. DNA sequencing revealed that she has harbored compound heterozygous variants of the F12 gene, namely c. 712_713insT (p.Cys238Leufs *73) in exon 8 and c. 1561G>A (p.Glu521Lys) in exon 13. Her mother and younger son were heterozygous for the p. Cys238Leufs*73 variant, while her older son was heterozygous for the p. Glu521Lys variant. Bioinformatic analysis suggested that Cys238 is highly conserved and p. Cys238Leufs*73 is a pathogenic variant, which eventually resulted in a truncated protein. Conclusion:The c. 712_713insT and c. 1561G>A compound heterozygous variants of the F12 gene probably underlay the decreased FⅫ level in this pedigree, among which c. 712_713insT (NM_000505) was unreported previously.

Hyperleukocytic actue leukaemia served as an acute and critical disease of the blood system,its fatality rate can be as high as 20%-30%,and the systemic treatment should be carried out immediately af-ter diagnosis.At present,a number of studies at home and abroad have confirmed that the patients with hyper-leukocytic acute leukemia can achieve a similar remission rate and prognosis as the patients with non-hyperleu-kocytic acute leukemia after effective leukocyte lowering pretreatment.This article discussed the aspects such as the mechanism of leukemia cytosis and leukocyte lowering pretreatment therapy,in order to provide a theoretical basis for the clinical diagnosis,treatment and research of this malignant hematological cancer.

OBJECTIVE: To analyze a Chinese pedigree with Hereditary coagulation factor Ⅻ (FⅫ) deficiency duo to variants of F12 gene and explore its molecular pathogenesis. METHODS: A patient who underwent laparoscopic cystectomy at the Department of Gynecology of the First Affiliated Hospital of Wenzhou Medical University in June 2012 was selected as the study subject. Coagulation factor indexes of the proband and her family members (5 individuals from three generations) were determined. All exons, flanking sequences, 5' and 3' untranslated regions of the F12 gene of the proband and her family members were analyzed by direct sequencing. Three bioinformatics software was used to analyze the conservation, pathogenicity and protein model of the variant. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No. 2012-17). RESULTS: The activated partial thromboplastin time (APTT), FⅫ activity (FⅫ:C) and FⅫ antigen (FⅫ:Ag) of the proband was 180.0 s, 1.0% and 2.1%, respectively. DNA sequencing revealed that she has harbored compound heterozygous variants of the F12 gene, namely c.712_713insT (p.Cys238Leufs *73) in exon 8 and c.1561G>A (p.Glu521Lys) in exon 13. Her mother and younger son were heterozygous for the p.Cys238Leufs*73 variant, while her older son was heterozygous for the p.Glu521Lys variant. Bioinformatic analysis suggested that Cys238 is highly conserved and p.Cys238Leufs*73 is a pathogenic variant, which eventually resulted in a truncated protein. CONCLUSION The c.712_713insT and c.1561G>A compound heterozygous variants of the F12 gene probably underlay the decreased FⅫ level in this pedigree, among which c.712_713insT (NM_000505) was unreported previously.
Adult ; Female ; Humans ; Male ; Middle Aged ; Base Sequence ; China ; Factor XII/genetics* ; Heterozygote ; Mutation ; Pedigree ; Factor XII Deficiency/genetics* ; East Asian People

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 a Chinese pedigree with Hereditary coagulation factor Ⅻ (FⅫ) deficiency duo to variants of F12 gene and explore its molecular pathogenesis. Methods:A patient who underwent laparoscopic cystectomy at the Department of Gynecology of the First Affiliated Hospital of Wenzhou Medical University in June 2012 was selected as the study subject. Coagulation factor indexes of the proband and her family members (5 individuals from three generations) were determined. All exons, flanking sequences, 5′ and 3′ untranslated regions of the F12 gene of the proband and her family members were analyzed by direct sequencing. Three bioinformatics software was used to analyze the conservation, pathogenicity and protein model of the variant. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No. 2012-17). Results:The activated partial thromboplastin time (APTT), FⅫ activity (FⅫ: C) and FⅫ antigen (FⅫ: Ag) of the proband was 180.0 s, 1.0% and 2.1%, respectively. DNA sequencing revealed that she has harbored compound heterozygous variants of the F12 gene, namely c. 712_713insT (p.Cys238Leufs *73) in exon 8 and c. 1561G>A (p.Glu521Lys) in exon 13. Her mother and younger son were heterozygous for the p. Cys238Leufs*73 variant, while her older son was heterozygous for the p. Glu521Lys variant. Bioinformatic analysis suggested that Cys238 is highly conserved and p. Cys238Leufs*73 is a pathogenic variant, which eventually resulted in a truncated protein. Conclusion:The c. 712_713insT and c. 1561G>A compound heterozygous variants of the F12 gene probably underlay the decreased FⅫ level in this pedigree, among which c. 712_713insT (NM_000505) was unreported previously.

Standards are the technical support for economic activities and social development. The construction and standardization of the pathogenic microorganism preservation standard system is an important technical foundation for the high-quality development of preservation work. Establishing a pathogenic microorganism resource standard system is also important to the national biosafety standards. Through the standardization of pathogenic microbial resource preservation, we can ensure the effective management and sustainable utilization of pathogenic microbial resources, promote the transformation of resources, and serve as an important new element of new productivity to assist the innovative development of biosafety science and technology. This article elaborates and analyzes the establishment background, construction framework, standardization process, and application effects of the standard system for preserving pathogenic microbial resources, providing stronger support for further improving the standard system and promoting the standardization of pathogenic microbial resource preservation.

Objective To investigate the inhibitory effects and mechanisms of α-hederin,an active ingredient in Fruc-tus Akebiae,on hepatocellular carcinoma(HCC)cells.Methods HCC cells were divided into four groups and treated with α-hederin(0,10,20,and 30 μmol·L-1)for 24 h and 48 h,respectively.MTT assays were used to detect the cell proliferation rate,flow cytometry(FCM)was used to detect the apoptotic rate,transcriptomics was used to screen signaling pathways in α-hederin-treated HCC cells,RNA interference was exploited to verify the underlying signaling pathway,and real-time quantitative PCR(qRT-PCR)and Western blotting(WB)were used to detect expression changes of the mRNA and protein of TP53(p53),PMAIP1(Noxa),and apoptosis-associated proteins,Caspase9 and Caspase3.Results α-Hederin induced apoptosis by activa-ting apoptosis-associated proteins,PARP,Caspase9 and Caspase3.Transcriptomics,qRT-PCR,and WB results also showed that α-hederin increased the mRNA and protein expression of p53 and Noxa.Furthermore,α-hederin inhibited the protein degradation of p53 and Noxa,reversing the apoptosis decrease in p53/Noxa siRNA-knocked-down HCC cells.In vivo results showed that α-hederin inhibited the growth of HCC tumors.Conclusion α-hederin may induce the apoptosis of HCC cells by activating and stabilizing the p53/Noxa signaling pathway.

In recent years,the"One Hospital with Multiple Campuses"model has emerged in major public hospitals across China,providing intrinsic impetus and vitality for the expansion and balanced distribution of high-quality medical re-sources.This paper analyzed the current situation and challenges of the multiple campuses development in public hospitals within China,adopting a problem-oriented approach to explore optimized path to a unified management system.The aim was to establish a collaborative management model across campuses based predicated on the homogenization of systems,management,supervi-sion,professionals,and performance.Taking the Third Affiliated Hospital of Sun Yat-sen University as a case in this study,the paper reviewed and summarized management strategies and practical experience from the aspects of management model,resource allocation,information-orientation,homogenized management,and cultural development.Additionally,the article suggested to u-tilize 5G technology to empower homogenization across campuses.In this way,the hospital's overall operational efficiency can be improved,optimize resource allocation optimized,homogenized medical services ensured,and core competitiveness enhanced.These efforts will provide a reference for the high-quality development of multi-campus public hospitals.