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 In order to shorten the transparency time of clear,unobstructed brain imaging cocktails and computational analysis(CUBIC),improve the transparency efficiency,and explore the possibility of applying hydrophilic tissue transparency technique,this study was conducted to optimize the perfusion of CUBIC technique and compare it with four hydrophilic tissue clearing method in terms of tissue transparency effect,transparency time,area change,volume change and adeno-associated virus(AAV)fluorescence retention.Methods Brain,liver,spleen and kidney of 6 adult Institute of Cancer Research(ICR)mice were subjected to clearing treatment by SeeDB,FRUIT,ScaleS and CUBIC method,respectively.The area and gray value of the samples were measured by Image J 1.8.0,and the volume before and after transparency was measured by drainage method to compare the transparency effect,time and size deformation of each group.Perfusion optimization of the CUBIC was performed by improving the perfusion rate with the optimal perfusion dose,each group of the experimental sample size was 6.Fluorescence preservation by different techniques was evaluated by injecting AAV in the motor cortex of 16 adult mice and taking the cervical spinal segments for transparency treatment after four weeks,and the fluorescence photographs were measured by Image J 1.8.0 to measure the mean fluorescent intensity.Results The optimal perfusion rate and dose of CUBIC was 15 ml/min and 200 ml respectively.For transparency ability and speed,the perfusion CUBIC had the lowest mean gray value and took the shortest time,while CUBIC consumed the longest time,and SeeDB,FRUIT,and ScaleS did not show good transparency ability.In terms of area and volume changes,several techniques showed different degrees of expansion after transparency of tissues or organs.In terms of fluorescence retention,perfusion CUBIC showed the best retention of green fluorescent protein(GFP)fluorescence signal,followed by CUBIC,ScaleS,FRUIT,and SeeDB.Conclusion Perfusion CUBIC technique shows the best tissue transparency,the shortest transparency time,and the most AAV fluorescence retention compared with other techniques.

The characteristic features of dissociative conversion disorder entail a partial or complete loss of normal integration in memory,identity awareness,immediate sensation,and motor control.This article documents a case of the disorder presenting with an onset of depressive mood,diminished interest,and facial tics,subsequently accompanied by intermittent fainting spells with limb convulsions.Despite multiple consultations in both general hospitals and psychiatric settings,and despite treatment with various antidepressants and antiepileptic medications,the patient's symptoms showed no significant improvement.The report of this case deepens the understanding of a complex and commonly misdiagnosed condition,offering valuable insights for the diagnosis and treatment of patients exhibiting similar symptoms.

AIM To optimize the processing technology of red Notoginseng Radix et Rhizoma and evaluate its blood tonifying activity.METHODS On the basis of a single factor experiment,with steaming temperature,steaming time,drying temperature,and drying time as influencing factors,the total contents of notoginsenoside R1,ginsenoside Rg1,Rb1,Rk3,Rh4,and 20(R)-ginsenoside Rg3 as evaluation indicators,Box-Behnken response surface method ology was used to optimize the processing technology.Upon the anemic mouse models jointly induced by 1-acetyl-2-phenylhydrazine(APH)and cyclophosphamide(CTX),the investigation of the blood tonifying activity of red Notoginseng Radix et Rhizoma was carried out in contrast to that of the steamed Notoginseng Radix et Rhizoma.RESULTS The optimal conditions,contributing saponin content of 8.326%and RSD of 0.087%,were determined as follows:steaming temperature of 130℃,steaming time of 4 hours,drying temperature of 60℃,and drying time of 48 h.The pharmacological activity revealed that the different processing techniques were responsible for the different blood enriching activity of notoginseng,with red Notoginseng Radix et Rhizoma displaying a better efficacy than that of steamed Notoginseng Radix et Rhizoma.CONCLUSION This stable and feasible method can be used to control the production of red Notoginseng Radix et Rhizoma.

Objective:To investigate the relationship between serum 25-hydroxyvitamin D [25(OH)D] and arterial stiffness in the physical examination population.Methods:It was a cross-sectional study, the data were collected from adults who received physical examination in the Health Management Center of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology from January 2016 to April 2022, all the participants completed the tested for brachial and ankle pulse wave conduction velocity (baPWV) and serum 25(OH)D. The general information, anthropometric metrics and relevant laboratory data were recorded. A total of 41 090 participants were included. According to baPWV, the subjects were divided into arterial stiffness group (baPWV≥1 800 cm/s, n=2 440) and normal group (baPWV<1 800 cm/s, n=38 650). The subjects were categorized into three groups based on serum 25(OH)D level: sufficient group [25(OH)D≥30 μg/L, n=2 830], insufficient group [25(OH)D 20-<30 μg/L, n=11 400] and deficiency group [25(OH)D<20 μg/L, n=26 860]. Logistic regression analysis was performed to assess the relationship between 25(OH)D and arterial stiffness. The dose-response relationship between serum 25(OH)D levels and the risk of arterial stiffness was analyzed using restricted cubic spline functions. Results:Of the 41 090 participants included, 25 001 were males and 16 089 were females, 2 440 cases (5.94%) were detected with arterial stiffness. Compared with normal group, the age, body mass index, uric acid, creatinine, 25(OH)D level and proportion of dyslipidemia were all significantly higher in the arterial stiffness group [(60.1±9.8) vs (46.0±10.4) years, (25.3±3.4) vs (24.5±3.4) kg/m 2, (356.2±99.6) vs (345.0±95.8) μmol/L, (77.0±21.5) vs (73.8±15.9) μmol/L, (18.51±7.93) vs (18.11±7.27) μg/L, 48.4% vs 38.5%], while the glomerular filtration rate was significantly lower [(87.2±16.1) vs (99.1±14.0) ml·min -1·1.73 m -2] (all P<0.05). The 25(OH)D deficiency was positively correlated with the risk of arteriosclerosis ( OR=1.26, 95% CI: 1.07-1.51), and the risk of arterial stiffness increased by 16% for every 10 μg/L decrease in serum 25(OH)D levels (all Ptrend<0.001). The mean baPWV level in the 25(OH)D deficiency group was 12.45 cm/s (95% CI: 3.89-21.02) higher than that in 25(OH)D adequacy group; and for every 10 μg/L decrease in 25(OH)D, the baPWV increased by 11.23 cm/s (95% CI: 8.11-14.19) (all Ptrend<0.001). The association between serum 25(OH)D deficiency and the risk of arteriosclerosis was more pronounced in individuals with a body mass index of 24 kg/m2 or above ( Pfor interaction=0.039). Moreover, there was an “L”-shaped dose-response relationship between serum 25(OH)D level and the risk of arteriosclerosis ( Pnon-linear<0.001). Conclusion:The 25(OH)D deficiency is closely correlated with the risk of arteriosclerosis in the physical examination population, and the association is stronger in overweight individuals.

Objective To investigate the risk factors for recurrence of persistent atrial fibrillation(AF)after radiofrequency ablation with circumferential pulmonary vein isolation+top line+back wall line based on the features of cardiac imaging and serum biomarkers,and then to establish a nomogram risk prediction model.Methods A total of 172 patients with persistent AF admitted to our hospital from June 2022 to September 2023 were enrolled and then according to recurrence or not in 6 months after surgery,they were divided into the recurrence group(51 cases)and the non-ecurrence group(121 cases).Before surgery,routine electrocardiography,and transthoracic and esophageal echocardiography were performed,while blood routine indicators and related biochemi-cal indicators were measured.All patients underwent radiofrequency ablation with circumferential pulmonary vein isolation,top line,and back wall line.They were followed up for 6 months after surgery.Binary logistic analysis was used to analyze the independent risk factors for postoperative recurrence,and then a nomogram risk prediction model was constructed and its diagnostic per-formance was evaluated.Results Lager LAD,higher LAVI,neutrophil count and NLR,and ele-vated BNP and CRP levels,while lower LAAFV,LAAEV and LAAEF were observed in the re-current group than those in the non-recurrent group(P＜0.01).Binary logistic regression analysis showed that elevated LAVI(OR=1.160,95％CI:1.006-1.337),decreased LAAEV(OR=0.740,95％CI:0.583-0.940),decreased LAAEF(OR=0.608,95％CI:0.422-0.877),elevated BNP(OR=1.017,95％CI:1.004-1.030),and higher NLR(OR=10.116,95％CI:1.316-77.755)were independent risk factors for recurrence after radiofrequency ablation of pulmonary vein isolation+top line+posterior wall line in persistent AF patients(P＜0.05,P＜0.01).The AUC value of the nomogram model constructed with LAVI,LAAEV,LAAEF,BNP and NLR in predicting postop-erative recurrence was 0.889(95％CI:0.833-0.932).Conclusion The cardiac imaging parame-ters LAVI,LAAEV and LAAEF,and serum biomarkers BNP and NLR are closely associated with postoperative recurrence of persistent AF in patients after radiofrequency ablation with cir-cumferential pulmonary vein isolation+top line+back wall line,and the relevant nomogram mod-el has better diagnostic value for postoperative recurrence.

Objective To develop and validate a deep learning model for automatic identification of liver CT contrast-enhanced phases.Methods A total of 766 patients with liver CT contrast-enhanced images were retrospectively collected.A three-phase classification model and an arterial phase(AP)classification model were developed,so as to automatically identify liver CT contrast-enhanced phases as early arterial phase(EAP)or late arterial phase(LAP),portal venous phase(PVP),and equilibrium phase(EP).In addition,221 patients with liver CT contrast-enhanced images in 5 different hospitals were used for external validation.The annotation results of radiologists were used as a reference standard to evaluate the model performances.Results In the external validation datasets,the accuracy in identifying each enhanced phase reached to 90.50%-99.70%.Conclusion The automatic identification model of liver CT contrast-enhanced phases based on residual network may provide an efficient,objective,and unified image quality control tool.

Establishing a new paradigm for high-quality development in public hospitals,embracing a comprehensive,systematic,and innovative strategic management philosophy,and instituting a modern hospital management system hold significant practical importance.It investigates the pathways through which the Guangzhou Women and Children's Medical Center has fulfilled its strategic commitments over its 17-year history.It delves into the meaning and value of constructing hospital strategic commitments,explores pathways for realization,analyzes effects.It aims to understand the normative role of strategic commitments in shaping the development paths of specialized women's and children's hospitals and to provide a reference for the high-quality development of similar hospitals across China.

Establishing a new paradigm for high-quality development in public hospitals,embracing a comprehensive,systematic,and innovative strategic management philosophy,and instituting a modern hospital management system hold significant practical importance.It investigates the pathways through which the Guangzhou Women and Children's Medical Center has fulfilled its strategic commitments over its 17-year history.It delves into the meaning and value of constructing hospital strategic commitments,explores pathways for realization,analyzes effects.It aims to understand the normative role of strategic commitments in shaping the development paths of specialized women's and children's hospitals and to provide a reference for the high-quality development of similar hospitals across China.

Establishing a new paradigm for high-quality development in public hospitals,embracing a comprehensive,systematic,and innovative strategic management philosophy,and instituting a modern hospital management system hold significant practical importance.It investigates the pathways through which the Guangzhou Women and Children's Medical Center has fulfilled its strategic commitments over its 17-year history.It delves into the meaning and value of constructing hospital strategic commitments,explores pathways for realization,analyzes effects.It aims to understand the normative role of strategic commitments in shaping the development paths of specialized women's and children's hospitals and to provide a reference for the high-quality development of similar hospitals across China.