Objective To investigate the antimicrobial resistance of clinical isolates from elderly patients(≥65 years)in major medical institutions across China.Methods Bacterial strains were isolated from elderly patients in 52 hospitals participating in the CHINET Antimicrobial Resistance Surveillance Program during the period from 2015 to 2021.Antimicrobial susceptibility test was carried out by disk diffusion method and automated systems according to the same CHINET protocol.The data were interpreted in accordance with the breakpoints recommended by the Clinical and Laboratory Standards Institute(CLSI)in 2021.Results A total of 514 715 nonduplicate clinical isolates were collected from elderly patients in 52 hospitals from January 1,2015 to December 31,2021.The number of isolates accounted for 34.3％of the total number of clinical isolates from all patients.Overall,21.8％of the 514 715 strains were gram-positive bacteria,and 78.2％were gram-negative bacteria.Majority(90.9％)of the strains were isolated from inpatients.About 42.9％of the strains were isolated from respiratory specimens,and 22.9％were isolated from urine.More than half(60.7％)of the strains were isolated from male patients,and 39.3％isolated from females.About 51.1％of the strains were isolated from patients aged 65-＜75 years.The prevalence of methicillin-resistant strains(MRSA)was 38.8％in 32 190 strains of Staphylococcus aureus.No vancomycin-or linezolid-resistant strains were found.The resistance rate of E.faecalis to most antibiotics was significantly lower than that of Enterococcus faecium,but a few vancomycin-resistant strains(0.2％,1.5％)and linezolid-resistant strains(3.4％,0.3％)were found in E.faecalis and E.faecium.The prevalence of penicillin-susceptible S.pneumoniae(PSSP),penicillin-intermediate S.pneumoniae(PISP),and penicillin-resistant S.pneumoniae(PRSP)was 94.3％,4.0％,and 1.7％in nonmeningitis S.pneumoniae isolates.The resistance rates of Klebsiella spp.(Klebsiella pneumoniae 93.2％)to imipenem and meropenem were 20.9％and 22.3％,respectively.Other Enterobacterales species were highly sensitive to carbapenem antibiotics.Only 1.7％-7.8％of other Enterobacterales strains were resistant to carbapenems.The resistance rates of Acinetobacter spp.(Acinetobacter baumannii 90.6％)to imipenem and meropenem were 68.4％and 70.6％respectively,while 28.5％and 24.3％of P.aeruginosa strains were resistant to imipenem and meropenem,respectively.Conclusions The number of clinical isolates from elderly patients is increasing year by year,especially in the 65-＜75 age group.Respiratory tract isolates were more prevalent in male elderly patients,and urinary tract isolates were more prevalent in female elderly patients.Klebsiella isolates were increasingly resistant to multiple antimicrobial agents,especially carbapenems.Antimicrobial resistance surveillance is helpful for accurate empirical antimicrobial therapy in elderly patients.

Objective To explore the deeper understanding of the pregnancy and delivery experience of three-child pregnant and matemal women,and to provide a basis for healthcare personnel to provide more systematic,safe,and targeted perinatal healthcare services and care measures for three-child pregnant and matemal women.Methods Purposive sampling method was used to select 17 cases of three-child pregnant and matemal women who were admitted and delivered in a tertiary level-A matemal and child healthcare hospital in Nanjing from August 2022 to June 2023 for semi-structured interviews,and Colaizzi 7-step process of analyzing,summarizing,and refining the themes was used.Results A total of 4 themes were extracted,including determination of willingness to become pregnant,perceived risks of childbirth,perceived benefits to themselves and their families,diversified support needs.Conclusion The establishment of pregnancy intention of three-child pregnant women is affected by many factors.Relevant departments should actively implement the supporting measures of the three-child birth policy;healthcare workers should strengthen perinatal healthcare services for three-child mothers to reduce the risk of giving birth,actively strengthen their sense of benefits related to pregnancy,and establish a whole process of support system to promote the health of mothers and infants.

Objective To explore the characteristics of high-frequency users of pediatric telemedicine outpatient services and analyze the factors influencing the frequency of use,providing theoretical support for the promotion and broader application of pediatric telemedicine outpatient services.Methods 197 parents of patients were randomly selected for telephone questionnaire survey by cross-sectional research method,and were divided into high frequency and low frequency users according to their frequency of use.Based on annual usage frequency,participants were divided into high-frequency and low-frequency groups.Mann-Whitney U test and x2 test or Fisher exact probability method were used for univariate analysis,and multiple logistic regression model was used to analyze the independent influencing fac-tors of the difference in the frequency of pediatric outpatient telemedicine use.Results A total of 165 questionnaires were completed.Among high-frequency users,the proportion of provinces outside Beijing was higher than that of low-frequency users,the overall knowledge level of information was better than that of low-frequency users,the overall satisfaction evaluation was better than that of low-frequency users,and the proportion of gastroenterology pa-tients was higher than that of low-frequency users.Conclusion Pediatric medical institutions need to strengthen the pub-licity of the platform,improve service quality,improve user experience by optimizing technical means,further pro-mote Internet outpatient services,and expand its application potential in children's health management.

AIM To study the chemical constituents from the sticks and leaves of Croton cascarilloides Raeusch.and their biological activities.METHODS The 95％ethanol extract from the sticks and leaves of C.cascarilloides was isolated and purified by MCI,silica gel,Sephadex LH-20 and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.LPS-induced NO RAW264.7 cell model induced by LPS was used to evaluate its anti-inflammatory activity in vitro.GES-1 injury model induced by taurocholic acid was used to screen the gastric mucosal protection activity.RESULTS Fourteen compounds were isolated and identified as bullatantriol(1),(-)-boscialin(2),(+)-dehydrovomifoliol(3),3-(hydroxylacetyl)-indole(4),pinoresinol(5),3,7-dimethyl-octa-1,7-diene-3,6-ol(6),(+)-syringaresinol(7),curcasinlignan B(8),cleomiscosin C(9),cleomiscosinD(10),2,6-dimethyl-octa-1,7-dien-3,6-diol(11),vanillin(12),vanillic acid(13),methyl vanillate(14).Compound 4 had certain anti-inflammatory activity,with IC50 values of 73.62 μmol/L.The protective rates of 25 μmol/L compounds 1-4,6,9-12 and 14 on gastric mucosal epithelial cells were 30.07％,34.18％,23.91％,30.92％,17.51％,19.69％,31.76％,22.46％,30.56％and 14.49％,respectively.CONCLUSION Compounds 1-14 are isolated from this plant for the first time.Compound 4 shows anti-inflammatory activity,1-4,6,9-12 and 14 show different degrees of gastric mucosal epithelial cell protective activity.

Objective To explore the diagnostic value of artificial intelligence(AI)in retinal vein occlusion(RVO).Methods The fundus images from 9,000 patients who visited our ophthalmology department and underwent non-mydriat-ic fundus photography from July 2016 to June 2022 were selected.Among these images,700 fundus images of patients with RVO were screened.These fundus images were processed using AI in collaboration with School of Electrical Engineering and Automation of Henan Polytechnic University.Besides,the state-of-the-art two-stage algorithms(the Faster-RCNN model),advanced one-stage detection algorithms(the YOLOV4 and YOLOV5 models),and a modified YOLOV5 model spe-cifically designed for RVO were employed in this study to process the collected RVO data.In addition,a comparative analy-sis of the results was conducted.Moreover,the sensitivity and specificity of the YOLOV5 model for the diagnosis of RVO were evaluated based on the receiver operating characteristic(ROC)curve.Results The Faster-RCNN model demon-strated the highest detection accuracy;however,its parameter scale was significantly larger than that of other models,and its frame rate was only 8 frames per second,which did not meet real-time detection requirements.The detection accuracy of the modified YOLOV5 model was only 3％lower than that of the Faster-RCNN model,but it had a significantly smaller parameter scale compared with the Faster-RCNN model,achieving a frame rate of 22 frames per second higher than the Faster-RCNN model.The area under the ROC curve(AUC)plotted based on the Faster-RCNN,YOLOV4,YOLOV5,and modified YOLOV5 models was 0.946(95％CI:0.933-0.959),0.794(95％CI:0.772-0.816),0.864(95％CI:0.845-0.884),and 0.930(95％CI:0.915-0.944),respectively.The sensitivity and specificity of the modified YOLOV5 model for the diagnosis of RVO were 87.0％and 98.9％,respectively.Conclusion The modified YOLOV5 model,with its high detection accuracy,smaller parameter scale,and high sensitivity and specificity for the diagnosis of RVO,can be consid-ered a new AI-assisted diagnostic method for RVO.

Objective To observe the difference of bone micro-structure in different regions of proximal femur,micro-CT scanning was performed on 30 proximal femur specimens to explain the mechanism of proximal femur fracture and to provide anatomical basis for prosthesis design.Methods Totally 30 intact proximal femur specimens were obtained from 60-80 year-old cadavers.Micro-CT scanning was used to measure the trabecular thickness(Tb.Th),trabecular number(Tb.N),trabecular space(Tb.Sp),connectivity(Conn)and bone mineral density(BMD)and other parameters in 7 regions of proximal femur,including proximal pressure trabecular(PPT),distal pressure trabecular(DPT),femoral head-neck junction(FHNJ),head and neck of femoral neck(HNFN),the base of femoral neck(BPFN),intertrochanteric line(IL)and greater trochanter(GT).Results The bone mineral density of IL and GT were higher than those of BPFN,FHNJ,DPT and PPT.The trabecular thickness of GT was the largest,followed by IL,BPFN and HNFN,and the smallest was FHNJ,DPT and PPT.The trabecular space of IL was larger than that of GT,and the data of both were larger than those of other parts,among which DPT and PPT were the smallest.The trabecular number of IL and GT were the smallest,BPFN,HNFN and FHNJ were larger,and DPT was the largest.The volume fraction of IL was the smallest,BPFN and HNFN were larger,DPT and PPT were the largest.Conclusion The bone density,trabecular thickness,bone volume,and total volume of GT and IL in the proximal femur of elderly patients are all relatively large,so the reason for the high incidence of fractures is not due to weak internal bone microstructure;The bone density,trabecular thickness,and trabecular gap at the proximal and distal ends of the vertical trabecular bone are relatively small.If it is necessary to perform core decompression for prosthesis filling at this location,the design should be conducive to the mechanical conduction of the prosthesis and the regeneration of surrounding bone tissue.

Objective To explore the mechanism by which the sanguis draconis flavones(SDF)regulates the reactive oxygen species(ROS)/thioredoxin-interacting protein(TXNIP)pathway to mediate cell pyroptosis and improve cerebral ischemia-reperfusion injury(CIRI)in rats.Methods The experimental rats were randomly divided into the control group(Ctrl),the CIRI group,the low-dose SDF group(SDF-L),the high-dose SDF group(SDF-H),and the SDF-H+ROS/TXNIP pathway activator,trimethylamine oxide(TMAO)group(SDF-H+TMAO).Among them,except for the control group,the remaining rats all needed to establish the CIRI rat model by the modified suture method.Zea Longa scoring was performed on rats from each group.ELISA was used to detect the levels of serum inflammatory factors interleukin(IL)-1β,IL-18 and oxidative stress-related factors superoxide dismutase(SOD),malondialdehyde(MDA),glutathione peroxidase(GSH-Px).Flow cytometry was used to measure the ROS levels.Cerebral edema was detected.Cerebral infarction was detected by 2,3,5-triphenyl tetrazolium chloride(TTC)staining.HE staining was used to detect the pathological changes of brain tissue.Immunohistochemistry was used to detect the expression of pyrolytic effector protein dermolin D(GSDMD).Western blotting was used to detect the expression of proteins related to the ROS/TXNIP pathway.Results Compared with the control group,a large area of cerebral infarctions were observed in the brain tissue of the CIRI group,accompanied by mild hemorrhage and obvious infiltration of inflammatory cells.Neuronal cells underwent degeneration and necrosis,with sparse and disordered arrangement.The phenomena of nuclear condensation and nucleolus lysis were obvious.The Zea Longa score,cerebral infarction volume,brain tissue water content,levels of IL-1β,IL-18,ROS,MDA,and the expressions of GSDMD,TXNIP,nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),apoptosis-related punctate protein(ASC),and Caspase-1 increased,while the activities of SOD and GSH-Px decreased(P＜0.05).Compared with the CIRI group,the pathological damage of brain tissues in the SDF-L group and the SDF-H group was significantly improved.The Zea Longa score,cerebral infarction volume,brain tissue water content,levels of IL-1β,IL-18,ROS,MDA,and the expressions of GSDMD,TXNIP,NLRP3,ASC,and Caspase-1 decreased.The activities of SOD and GSH-Px increased(P＜0.05);TMAO treatment partially reversed the improvement effect of SDF on CIRI in rats.Conclusion SDF ameliorates cerebral CIRI in rats by inhibiting ROS/TXNIP pathway-mediated pyroptosis.

Objective To explore the effects of pretreatment with rehmannia glutinosa polysaccharide(RGP)and basic fibroblast growth factor(FGF-2)alone or in combination on the differentiation of bone marrow mesenchymal stem cells(BMSCs)into cardiomyoid cells and the related mechanisms.Methods BMSCs pretreated with FGF-2 and RGP alone or in combination were cultured for 1,2,and 4 weeks.The expression levels of NKx2.5 and GATA-4 were detected by Real-time PCR.BMSCs pretreated with FGF-2 or RGP alone and PI3K/Akt signaling pathway inhibitor LY294002 were used to detect the expression levels of myocardial specific proteins and related pathway proteins by Western blotting.Results Compared with the blank control group,pretreatment with FGF-2 or RGP alone increased the expression rate of myocardial specific markers,and the effect of combined pretreatment with FGF-2 and RGP was more obvious(P＜0.05).Compared with pretreatment with FGF-2 or RGP alone,pretreatment with LY294002 combined with FGF-2 or RGP significantly down-regulated the expression of cardiac-specific proteins in BMSCs and inhibited the phosphorylation of Akt(P＜0.05).Conclusion Both FGF-2 and RGP can induce BMSCs to differentiate into cardiomyocyte-like cells by regulating PI3K/Akt signaling pathway,and the combination of FGF-2 and RGP has a better inductive effect.

The CRISPR/dCas9 system is a gene editing tool that has proven to be highly efficient and precise. By utilizing transcriptional activators, such as VP64, p65, and Rta, the system can effectively and stably activate target genes. Sox17, a transcription factor belonging to the SOX family, plays a crucial role in the differentiation of the germ layers and the determination of cell fates during the early stages of embryonic development. Sheep embryonic stem cells (sESCs) are characterized by their capacity for self-renewal and multidirectional differentiation, serving as a significant in vitro model for studying the mechanisms of cell differentiation during early embryonic development. However, the importing of exogenous genes into sESCs is challenging due to their unique growth characteristics. The objective of this study was to investigate the conditions necessary for successfully activating Sox17 in sESCs. To this end, we employed the CRISPR/dCas9 system along with liposome transfection, lentivirus invasion, and electroporation to activate Sox17 in sESCs. The expression of Sox17 was then determined by fluorescence quantitative PCR, on the basis of which the performance of different transfection methods was compared. The results indicated that the electroporation group had the best transfection effect and the highest Sox17 expression among the three transfection methods. The efficient and stable gene activation protocol will provide a reference for embryonic stem cell research in other species, especially livestock animals, and lay the foundation for the subsequent study of gene function and realization of precise cell fate regulation by regulating gene expression in sheep embryonic stem cells.
Animals ; CRISPR-Cas Systems/genetics* ; Sheep ; SOXF Transcription Factors/genetics* ; Embryonic Stem Cells/cytology* ; Genetic Vectors/genetics* ; Cell Differentiation/genetics* ; Transfection ; Gene Editing/methods*

Objective:To provide guidance on parameters for future endovaginal coil magnetic resonance imaging(MRI)in vivo,and to investigate the imaging potential of the endovaginal coil technique.Methods:The endovaginal coil was used to perform T2-weighted turbo spin echo sequence imaging on a phantom containing three spatial resolution modules,with variations in key imaging parameters.Signal-to-noise ratio(SNR),contrast-to-noise ratio(CNR),and contrast(C)were calculated for images under different imaging parameters,and image quality was assessed based on scores within specified intervals.Structural display scores were determined for the three resolution modules of the phantom,and the non-parametric tests were used for statistical comparison between groups.Results:Under the main imaging parameters of a slice thickness of 2 mm,a slice gap of 0.2 mm,4 times of excitation repetition,a matrix size of 384×384,an FOV of 52 mm×52 mm,a TE of 74 ms,a bandwidth of 180 Hz/Pixel,and an acceleration factor of 18,good image quality was obtained with an acquired spatial resolution of 0.14 mm×0.14 mm×2.00 mm and a reconstructed resolution of 0.07 mm×0.07 mm×2.00 mm,and all three resolution modules were clearly visualized with a clinically acceptable imaging time.There were signifi-cant differences between groups in the imaging quality scores of slice thickness,slice gap,number of excitations,FOV,and accelera-tion factor(P<0.05),while there were no significant differences in matrix,TE,and bandwidth between groups(P=0.521,0.502,and 0.566).There were significant differences in the structural display scores for all parameters between groups(P<0.05).Conclusion:The parameters obtained in this experiment can guide the future ap-plication of endovaginal coils in the human body,demonstrating the extremely high imaging potential of this technique,and endovaginal coil has the potential of identifying small lesions in future clinical practice.