Objective To evaluate the feasibility and effectiveness of four closure techniques ,large incision with plastic distal at‐tachment and clip with suction ,1 .1 cm small incision with plastic distal attachment and clip with suction in natural orifice translu‐minal endoscopic surgery(NOTES) .Methods Forty‐one ex vivo porcine stomachs were involved in this research .According to the size of incision and different methods of incision closure ,all ex vivo porcine stomachs were divided into four groups .Group A in‐volved 8 ex vivo porcine stomachs ,their size of incision were 1 .2-2 .0 cm ,and their incisions were closured by a clip closure direct‐ly ;group B involved 10 ex vivo porcine stomachs ,their size of incision were 1 .2-2 .0 cm ,and their incisions were closed by plastic end attachment with suction and clip enclosure technique;group C involved 10 ex vivo porcine stomachs ,their size of incision were smaller than 1 .1 cm ,and their incisions were by a clip closure directly ;group D involved 13 ex vivo porcine stomachs ,their size of incision were smaller than 1 .1 cm ,and closured by plastic end attachment with suction and clip enclosure technique .Stomach leaks were evaluated by leaking studies after the procedure .Results In group A ,1 incision was closed successfully ,7 incisions were clo‐sured unsuccessfully ;in group B ,2 incisions were closed successfully ,8 incisions were closed unsuccessfully ;in group C ,2 incisions were closed successfully ,8 incisions were closured unsuccessfully ;in group D ,11 incisions were closed successfully ,2 incisions were Closured unsuccessfully .Fisher′s exact test showed that group D was significantly related to the success of incision closure .The were significant differences between group A and D ,group B and group D(P<0 .05) .Conclusion Small incision(smaller than 1 cm) and plastic end attachment with suction and clip enclosure technique are optimal in NOTES procedure in this ex vivo porcine stomachs study .

As a safety culture placed between a culture of punishment and that of exemption, just culture can effectively improve the reporting rate of medical adverse events, promote medical staff to learn from events, and continuously improve patient safety management. The authors searched for literature on the study of just culture in medical institutions both domestically and internationally, including such databases as CNKI, Wanfang Data, and PubMed from 2001 to January 2023. They reviewed the concept of just culture and its connotation in the field of patient safety management, as well as the current research status of just culture in medical institutions both domestically and internationally. On such basis, they proposed for the construction of just culture in medical institutions in China, for reference in the construction and evaluation of a just culture in such institutions.

OBJECTIVEMutations of presenilin 1 (PSEN1) gene are the most frequent cause for familial Alzheimers disease (AD). This study was set to explore potential mutation of PSEN1 gene in a Chinese family featuring early-onset Alzheimers disease (FAD).

METHODSDNA was isolated from peripheral blood samples from 17 members of the FAD family as well as 10 patients with sporadic Alzheimers disease and 100 healthy subjects. With polymerase chain reaction (PCR) and Sanger sequencing, exons 113 of the PSEN1 gene were analyzed.

RESULTSDNA sequencing has revealed a heterozygous point mutation from G to A at position 1133 (Gly378Glu) of exon 11 of PSEN1 gene in 6 members from the family, among whom 5 were patients with dementia, whilst the remaining 1 was clinically normal but under onset age. The same mutation was not found in all other patients and the normal controls.

CONCLUSIONA novel missense mutation of the PSEN1 gene, Gly378Glu, probably underlies the autosomal dominant early-onset FAD in this Chinese family.

Adult ; Aged ; Alzheimer Disease ; diagnosis ; genetics ; Base Sequence ; Female ; Humans ; Male ; Middle Aged ; Pedigree ; Presenilin-1 ; genetics

Objective The optimal access for natural orifice transluminal endoscopic surgery is still uncertain .This study was designed to compare the practicability and maneuverability of transgastric ,transunmbilical ,and transrectal approach in abdominal surgery in a canine model .Methods Three dogs were used in this research .Three approach :trangastric ,transunmbilical and tran-srectal approach were carried out for abdominal exploration ,liver biopsy ,bladder biopsy and an attempted cholecystectomy .The ma-neuverability ,endoscopic image ,performer′s perception ,and spatial orientation were evaluated .Results The maneuverability of trangastric ,and transrectal approach NOTES were better than transunmbilical NOTES .Abdominal exploration ,live biopsy ,and bladder biopsy were completed successfully .The cholecystectomy was failed because of poor exposure and difficulty of separating the around tissure .Conclusion The optimal approach for upper abdomen NOTES is transrectal route .For lower abdomen NOTES , the trangastric approach is superior to other accesses .Further study is needed to develop more flexible and precise equipment for NOTES and to evaluate more feasible access approach .

Objective: To identify the functional connectivity characteristics of insula, sensory and social related brain regions in boys with autism spectrum disorder(ASD), to explore the central nervous basis of sensory abnormality affecting core symptoms in boys with ASD. Methods: Resting state functional magnetic resonance imaging (rs fMRI) data were collected from 34 boys with ASD and 29 typical development boys (TD group). Based on functional connectivity analysis, the sensory related brain regions, insula, and social related brain regions were taken as regions of interest to calculate the functional connectivity (FC) level between the regions of interest, the differences between the two groups were compared and the results were corrected by FDR. The Autism Diagnostic Observation Schedule (ADOS), Childhood Autism Rating Scale (CARS) and Autism Spectrum Quotient-Children s Version (AQ-Child) were used to assess the core phenotypes of boys with ASD. Results: Compared with the TD group, the levels of FC between tactile brain regions and insula, olfactory brain regions and insula, auditory brain regions and insula in boys with ASD group were significantly increased. The level of FC between the insula and bilateral amygdala，insula and the medial prefrontal cortex(mPFC) were significantly increased( P <0.05).Pearson correlation analysis showed that the level of FC between auditory brain region(BA42)and left insula in ASD group was negatively correlated with the scores of communication subscale of ADOS( r =-0.44)，social interaction subscale of ADOS( r =-0.43), communication & social interaction subscale of ADOS( r =-0.49),attention to details subscale of AQ-Child( r =-0.41). The level of FC between the right insula and right amygdala was positively correlated with the attention switch subscale of AQ-Child( r =0.38), the level of FC between right insula and mPFC was positively correlated with the scores of repetitive behavior subscale of ADOS( r =0.48), the attention switch subscale of AQ-Child( r =0.49), total scale subscale of AQ-Child( r =0.41), total scale of CARS( r =0.41)( P < 0.05 ). Conclusion The levels of FC between insula and sensory related, social related brain regions are abnormal in children with ASD, which have significant correlations with clinical symptoms. In depth studies can be conducted to explore underlying neutral mechanisms.

OBJECTIVE: To explore the genetic etiology for a Chinese pedigree affected with Angelman syndrome (AS). METHODS: The proband with phenotypes suggestive of AS was subjected to copy number variation sequencing (CNV-seq), methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) and high-throughput next generation sequencing (NGS). Variant of the UBE3A gene was verified among family members by Sanger sequencing and bioinformatic analysis. RESULTS: NGS revealed that the proband has carried a heterozygous variant of the UBE3A gene, namely c.1517G>A (p.R506H). The variant has co-segregated with the disease in the pedigree. Multiple amino acid sequence alignment showed that the site of mutant residue is conserved among nine homologous species. The variant was predicted to be deleterious by bioinformatic analysis. CONCLUSION A novel variant of the UBE3A gene has been identified in a Chinese pedigree affected with AS. Above finding has further expanded the spectrum of UBE3A gene variants and phenotypes of AS, which also facilitated molecular diagnosis and genetic counseling for the family.
Angelman Syndrome/genetics* ; China ; DNA Copy Number Variations ; Humans ; Mutation ; Pedigree ; Phenotype

Objective ThispaperpresentedaninvestigationonthesimilaritiesanddifferencesintheclinicalfeaturesandCTfindings betweenallogeneichematopoieticstem celltransplantation (allo-HSCT)inducedandnon-transplantinducedair-leaksyndromes (ALS)inpatientssufferingfromhematopathy,toimprovetheunderstandingofALSinpatientswithhematopathy.Methods Retrospective analysesandcomparisonsofclinicaldataandCTimageswereconductedbetweenGroupA (12patientswithALSafterallo-HSCT) andGroupB (26patientswithnon-transplant-relatedALS).A M annG W hitney U testwasperformedtoevaluatethemeasurementdata, andthe χ 2testor Fisher exacttestwasconductedtoexaminetheenumerationdata.Differencethresholdsof P＜0.05from bothsides weretakentobethedeterminantforstatisticalsignificance.Results TheincidenceratesofALSinpatientswithhematopathyafter anallo-HSCTwerefoundtobesignificantlyhigherthanthoseinpatientsonwhomsuchtransplantshadnotbeenperformed(1.84%. vs.0.06%),P＜0.001.SymptomsofdyspneaweremuchmorefrequentlyobservedingroupAcomparedtogroupB (7/12vs1/26), P＜0.01;whereasthedifferencesforthesymptomsofchesttightness,chestpain,andpharyngalgia werenotadequateintermsofstatistical significance,P＞0.05.IngroupA,theoccurrencesofALSsecondarytolongonsetnon-infectionpulmonarycomplications(LONIPC) associatedwithchronicgraft-versus-hostdisease(cGVHD)werefoundin8/12patients,whereastheoccurrencesin15/26patients weresecondarytopulmonaryinfectioningroupB,P＜0.01.Therewerenostatisticallysignificantdifferencesinage,gender,BMI, backgroundblooddisease,basictreatmentcounts,CTtype,treatmentmethodsandCTdisappearancetimelengthbetweenthetwo groups,P＞0.05.Conclusion Thereweredifferencesintheincidencerates,basiclungdiseasesandclinicalsymptomsbetweenallo-HSCT inducedandnon-transplantinducedALSinpatientssufferingfromhematopathy.Hematopathy-associatedALSwascommoninyoung adultswithlankypostures,patientswithleukemiaasback-grounddisease,patientswithahistoryofchemotherapyandpatientswith pulmonarydiseases.Thecommonsymptomsofpatients with hematopathy-associated ALS were chesttightness and chest pain,andpatients’overallprognosisweregood,meanwhileCT manifestationsweremainlycharacterizedbymixedpulmonary interstitialemphysema(PIE)+pneumomediastinum (PM)andsimplepneumothorax (PT).

The antigen gene expression level of a DNA vaccine is the key factor influencing the efficacy of the DNA vaccine. Accordingly, one of the ways to improve the antigen gene expression level of a DNA vaccine is to utilize a plasmid vector that is replicable in eukaryotic cells. A replicative DNA vaccine vector pCMVori was constructed based on the non-replicative pcDNA3.1 and the replicon of porcine circovirus 2 (PCV2) in this study. An EGFP gene was cloned into pCMVori and the control plasmid pcDNA3.1. The two recombinant vectors were transfected into PK-15 cell, and the plasmid DNA and RNA were extracted from the transfected cells. Real-time PCR was used to determine the plasmid replication efficiency of the two plasmids using plasmid before and after Bcl Ⅰ digestion as templates, and the transcription level of the Rep gene in PCV2 replicon was detected by RT-PCR. The average fluorescence intensity of cells transfected with the two plasmids was analyzed with software Image J, and the transcription level of EGFP was determined by means of real-time RT-PCR. The results showed that the replication efficiency of pCMVori in PK-15 cells incubated for 48 h was 136%, and the transcriptions of Rep and Rep' were verified by RT-PCR. The average fluorescence intensity of the cells transfected with pCMVori-EGFP was 39.14% higher than that of pcDNA3.1-EGFP, and the transcription level of EGFP in the former was also 40% higher than that in the latter. In conclusion, the DNA vaccine vector pCMVori constructed in this study can independently replicate in eukaryotic cells. As a result, the expression level of cloned target gene was elevated, providing a basis for developing the pCMVori-based DNA vaccine.
Animals ; Swine ; Circovirus/genetics* ; Vaccines, DNA/genetics* ; Replicon/genetics* ; Genetic Vectors/genetics* ; Plasmids/genetics*

Obesity is a worldwide epidemic. Promoting browning of white adipose tissue (WAT) contributes to increased energy expenditure and hence counteracts obesity. Here we show that cordycepin (Cpn), a natural derivative of adenosine, increases energy expenditure, inhibits weight gain, improves metabolic profile and glucose tolerance, decreases WAT mass and adipocyte size, and enhances cold tolerance in normal and high-fat diet-fed mice. Cpn markedly increases the surface temperature around the inguinal WAT and turns the inguinal fat browner. Further investigations show that Cpn induces the development of brown-like adipocytes in inguinal and, to a less degree, epididymal WAT depots. Cpn also increases the expression of uncoupling protein 1 (UCP1) and other thermogenic genes in WAT and 3T3-L1 differentiated adipocytes, in which AMP-activated protein kinase (AMPK) plays an important role. Our results provide novel insights into the function of Cpn in regulating energy balance, and suggest a potential utility of Cpn in the treatment of obesity.

ObjectiveThe type 2C protein phosphatases （PP2C） are involved in numerous plant signal transduction pathways. They mainly participate in plant stress response and regulate second metabolites biosynthesis via negatively regulating MAPK signaling pathway. Herein，we were to identify and analyze PP2C （CsPP2C） gene family from hemp genome，in hope of providing comprehensive insights for studying CsPP2C function during the development of hemp. MethodMolecular Evolutionary Genetics Analysis （MAGA）-X was used to construct phylogenetic tree. Expert Protein Analysis System （ExPASy），WoLF PSORT，Multiple EM for Motif Elicitation （MEME），Batch Conserved Domain Search （Batch-CD-Search），PlantCare，and TBtools were used，respectively，to predict CsPP2C physicochemical properties，subcellular localization，conserved motifs，protein structure，cis-element in promoter and collinearity with Arabidopsis PP2C. Cannabis sativa transcriptome and Real-time polymerase chain reaction（Real-time PCR） were used to analyze and verify gene expressions，respectively. ResultFifty-two CsPP2C with conserved domains were identified from the entire genome of hemp，encoding proteins ranging from 244 to 1 089 aa in length and with molecular weights ranging from 26.76 to 122.53 kDa. Those genes were mainly distributed in the nucleus，cytoplasm and chloroplast. The 47 CsPP2C were divided into 10 subfamilies，and the remaining 5 were not clustered. Seven pairs of homologous genes between hemp and Arabidopsis thaliana were identified according to collinear analysis. The light-responsive elements and abscisic acid elements are most abundant in the prediction. The gene expression heat map showed varied expression pattern of CsPP2C in different tissues. Real-time PCR results of three CsPP2C were consistent with transcriptome data. Moreover，alternative splicing analysis showed that some CsPP2C had alternative-splicing genes during evolution. ConclusionWe predicted and analyzed CsPP2C gene family in genomic scale and showed that CsPP2C are involved in many biological processes，whereby provides foundation for CsPP2C functional study.