Background::The ratio of high-density lipoprotein cholesterol (HDL-C) to low-density lipoprotein cholesterol (LDL-C) predicts cardiovascular disease (CVD) endpoints, yet its prognostic validity in high-risk populations and for type 2 diabetes mellitus (T2DM)-related adverse events remains unestablished.Methods::This study included 32,609 people aged 35-75 years in Fujian Province, China, who were at high risk for CVD. The primary endpoint was all-cause mortality during follow-up. Cox proportional hazard models and restricted cubic spline (RCS) analysis were used to evaluate the correlation between the HDL-C/LDL-C ratio and the endpoints.Result::On the basis of the restricted RCS curve, the participants were classified as having a low (< 0.3), middle (0.3-0.5), or high (> 0.5) HDL-C/LDL-C ratio. Multivariate Cox regression analyses revealed that the risk of all-cause mortality (HR = 1.48, 95% CI 1.14-1.93, p < 0.01 for low; HR = 1.30, 95% CI 1.06-1.58, p < 0.05 for high) was increased in the low and high groups. Participants without T2DM who were at high risk for CVD had similar prognoses (HR = 1.65, 95% CI 1.19-2.28, p < 0.01 for low; HR = 1.35, 95% CI 1.05-1.74, p < 0.01 for high). However, this association was not found in participants with T2DM who were at high risk for CVD. Conclusion::HDL-C/LDL-C can be used to predict the prognosis of individuals at high risk for CVD, and maintaining HDL-C/LDL-C ratios between 0.3 and 0.5 may be the most helpful range for this population. Furthermore, maintaining this ratio range holds clinical significance for cohorts without T2DM, although further exploration is needed in this T2DM cohort.

A VLi-net based cell nucleus segmentation method integrating convolutional neural networks(CNN)and Vision Transformer(ViT)is proposed to address the limitation that the U-Net with CNN as its backbone is only proficient in capturing local features and has a restricted receptive field.Firstly,to mitigate challenges such as high cost of data annotation and insufficient annotated data,text annotations are introduced to enhance the network's understanding of image information.Secondly,to improve the segmentation performance of VLi-net,ViT and CNN are combined to fully extract global and local features,with multi-receptive field convolution features incorporating into the ViT structure for effectively mitigating the issues of limited local information interaction and single feature representation in ViT.Finally,an interactive fusion module(ViFusion)is used to efficiently fuse the multi-level features from the CNN and ViT branches.Experimental results show that VLi-net achieves a Dice coefficient of 80.85%and a mean intersection over union(MIoU)of 66.83%on the MoNuSeg dataset,obtains a Dice coefficient of 80.53%and a MIoU of 67.54%on the DSB-2018 dataset,and has a Dice coefficient of 86.87%and a MIoU of 77.44%on the TNBC dataset.These findings confirm that VLi-net outperforms other methods across multiple experimental metrics.

Objective:To investigate the drug resistance gene characteristics, plasmid characteristics and genome tracing of Shigella sonnei causing a bacillary dysentery outbreak in Shandong Province. Methods:Sixty-five Shigella sonnei strains isolated from a 2021 outbreak in a county of Shandong Province were analyzed using antimicrobial susceptibility testing, whole genome sequencing (WGS), characterization of resistance and virulence genes, plasmid profiling, core genome multilocus sequence typing (cgMLST), and single nucleotide polymorphism (SNP) analysis. Results:All isolates had the same resistance phenotype and genotypes and were multidrug-resistant ESBL-producing Shigella sonnei, carrying important virulence genes. Plasmid analysis revealed a conserved genetic arrangement, pil( M/ N/ O2/ P)-tra( F/ H/ J/ K/ N/ O/ P/ Q)-IS Ecp1- blaCTX-M-14-Tn 903- yub( J/ I/ F/ G/ E/ D), and shared across strains from diverse regions and bacterial species. The cgMLST and SNP analyses demonstrated concordant clustering, with all 65 outbreak-related strains forming a single cluster alongside human-derived strains from Guangxi. Conclusion:The ESBL-producing Shigella sonnei responsible for the outbreak shares a homologous relationship with Guangxi human-derived strains, and the detected resistance plasmids and virulence genes underscore the need to strengthen drug resistance surveillance and genome tracing.

AIM:This study aims to investigate whether vitamin E succinate(VES)induces autophagy in hu-man gastric cancer cells through the promotion of mitochondria-associated endoplasmic reticulum membranes(MAMs).METHODS:Human gastric cancer cell lines MKN28 and MKN45 were cultured in vitro.Cell viability was assessed us-ing the CCK8 assay,and two cell growth curves were plotted to determine the treatment concentration of VES.Control groups,VES dose groups(MKN28:5,10,20,and 40 mg/L;MKN45:10,20,40,and 80 mg/L),an autophagy-posi-tive control group(rapamycin,RAPA,100 nmol/L),and a MAMs-positive control group(oligomycin A,10 mg/L)were set up.Cells were harvested after 24 h of treatment for subsequent experiments.The formation of autophagosomes and MAMs was observed using transmission electron microscopy.The expression levels of autophagy-related proteins,includ-ing beclin-1,LC3-II/LC3-I,and p62,were detected by Western blot.MAMs labeled with split green fluorescent protein(GFP)were visualized by fluorescence microscopy.The expression of mitofusin 2(MFN2),a key molecule of MAMs,was also detected by Western blot.To inhibit MFN2 specifically,the cells were treated with mitochondrial fusion inhibitor 8(MFI8)and simultaneously transfected with an MFN2 plasmid to achieve MFN2 overexpression(OE-MFN2).The cells were divided into control group,MFI8(20 μmol/L)group,VES groups(20 mg/L for MKN28 cells and 40 mg/L for MKN45 cells),VES+MFI8 group,OE-MFN2+MFI8 group and OE-MFN2+VES+MFI8 group.The MAMs were visualized by fluorescence microscopy,and the expression changes of MFN2,beclin-1 and LC3-II/I were detected by Western blot.RESULTS:The results of the CCK8 assay showed that VES significantly inhibited the viability of both human gastric can-cer cell lines(P<0.05).After VES treatment,the formation of typical autophagosomes and MAMs was observed in both cell lines by transmission electron microscopy.Fluorescence microscopy showed a significant increase in GFP signals of MAMs.Western blot analysis showed that with increasing doses of VES,the expression levels of MFN2,beclin-1,and LC3-II/I were significantly up-regulated,while that of p62 was significantly down-regulated(P<0.05).Compared with VES group,the cells pretreated with MFI8 followed by VES exposure showed markedly reduced GFP signals of MAMs and much lower protein levels of MFN2,beclin-1 and LC3-II/LC3-I(P<0.05).Transfection with an MFN2 overexpression plasmid rescued MFN2 expression.Compared with VES+MFI8 group,the cells in OE-MFN2+VES+MFI8 group had much higher protein expression levels of MFN2,beclin-1 and LC3-II/LC3-I(P<0.05).CONCLUSION:The VES may partici-pates in the regulation of autophagy in human gastric cancer cells by promoting the formation of MAMs.

This study aimed to investigate the effects of sweet potato leaf extract on production per-formance,systematic and mammary gland oxidative stress status and rumen microbiota of dairy goats fed high concentrate diets.Twenty Guanzhong dairy goats with same parity,similar lactation period(120±15)d and healthy body condition were selected and randomly divided into four groups:low-concentrate(LC),low-concentrate supplemented with 1％sweet potato leaf extract(LCS),high-concentrate(HC)and high-concentrate supplemented with 1％sweet potato leaf ex-tract(HCS).The experimental period was 35 days.The results showed that in the third week,milk yield in the HCS group was significantly higher than that in the LC and LCS groups(P＜0.05).The content of lipopolysaccharide in the rumen fluid of the HC group was significantly higher than that of the other three groups(P＜0.05),the content of malondialdehyde in the serum of the HC group was significantly higher than that of the LCS group(P＜0.05),the content of reactive oxygen species,protein carbonyl,8-hydroxydeoxyguanosine in the milk of the HC group was sig-nificantly higher than that of the LC and LCS groups(P＜0.05),GSH-Px in HCS group was sig-nificantly higher than that in the other three groups(P＜0.05).After the addition of sweet potato leaf extract,there was an increasing trend in the content of Anabaena phylum at the phylum level.In the joint analysis of genera,rumen fluid LPS showed highly significant negative correlation with Succiniclasticum(P＜0.01)and negative correlation with Prevotella(P＜0.05).Valeric acid was negatively correlated with Prevotella(P＜0.05).The pH value was negatively correlated with Treponema(P＜0.05).Butyric acid was positively correlated with Anaeroplasma(P＜0.05).In conclusion,the addition of sweet potato leaf extract to the diet can increase milk production and al-leviate the state of mammary gland oxidative stress,as well as improving rumen microbial diversi-ty of dairy goats.

This study aimed to investigate the effects of sweet potato leaf extract on production per-formance,systematic and mammary gland oxidative stress status and rumen microbiota of dairy goats fed high concentrate diets.Twenty Guanzhong dairy goats with same parity,similar lactation period(120±15)d and healthy body condition were selected and randomly divided into four groups:low-concentrate(LC),low-concentrate supplemented with 1％sweet potato leaf extract(LCS),high-concentrate(HC)and high-concentrate supplemented with 1％sweet potato leaf ex-tract(HCS).The experimental period was 35 days.The results showed that in the third week,milk yield in the HCS group was significantly higher than that in the LC and LCS groups(P＜0.05).The content of lipopolysaccharide in the rumen fluid of the HC group was significantly higher than that of the other three groups(P＜0.05),the content of malondialdehyde in the serum of the HC group was significantly higher than that of the LCS group(P＜0.05),the content of reactive oxygen species,protein carbonyl,8-hydroxydeoxyguanosine in the milk of the HC group was sig-nificantly higher than that of the LC and LCS groups(P＜0.05),GSH-Px in HCS group was sig-nificantly higher than that in the other three groups(P＜0.05).After the addition of sweet potato leaf extract,there was an increasing trend in the content of Anabaena phylum at the phylum level.In the joint analysis of genera,rumen fluid LPS showed highly significant negative correlation with Succiniclasticum(P＜0.01)and negative correlation with Prevotella(P＜0.05).Valeric acid was negatively correlated with Prevotella(P＜0.05).The pH value was negatively correlated with Treponema(P＜0.05).Butyric acid was positively correlated with Anaeroplasma(P＜0.05).In conclusion,the addition of sweet potato leaf extract to the diet can increase milk production and al-leviate the state of mammary gland oxidative stress,as well as improving rumen microbial diversi-ty of dairy goats.

Objective To propose a classification model based on a pseudo-bag strategy and feature adjustment for whole slide imaging in pathology.Methods A pseudo-bag generator was constructed to divide a parent bag into 3 pseudo-bags for increasing the number of training bags.Then,a pseudo-bag learning method based on Nystr?m-based algorithm for approximating self-attention and a selective feature fusion method were employed to process the pseudo-bags.Specifically,the pseudo-bag learning method based on Nystr?m-based algorithm for approximating self-attention reduced computational complexity through an improved multi-head self-attention mechanism while deeply extracting instance features to obtain pseudo-bag classification predictions,thereby enhancing pseudo-bag classification accuracy;and the selective feature fusion method refined pseudo-bag features by filtering and extracting relevant instances.Finally,the model adjusted bag features by extracting confounding factors to avoid interference from irrelevant information and further improve classification accuracy.Results The proposed model was evaluated on two datasets(CAMELYON-16 and TCGA-NSCLC)and compared with 10 other methods,and the results demonstrated that the proposed model achieved the best performance.The proposed method reached an accuracy of 0.943 on the CAMELYON-16 dataset and 0.906 on the TCGA-NSCLC dataset.Conclusion The proposed model can significantly improve the accuracy of whole-slide pathological image classification by effectively mitigating the overfitting and avoiding interference from irrelevant information.

The catalytic activity of 3-mercaptopyruvate (3MP) sulfurtransferase (MPST) converts 3MP to hydrogen sulfide (H₂S). However, the regulatory mechanisms governing MPST and its impact on the brain remain largely unexplored. Our study reveals the neuroprotective role of endothelial MPST-generated H₂S, regulated by protein phosphatase 2A (PP2A). Bioinformatics analysis and RNA sequencing demonstrated that endothelial PP2A is associated with neurodegenerative disease pathways. Cerebral ischemic mice exhibited significant inactivation of endothelial PP2A, evidenced by the reduction of PP2Acα in the brain endothelium. Mice with endothelium-specific null PP2A (PP2A^EC-cKO) exhibited neuronal loss, cognitive dysfunction, and long-term potentiation deficits. Postnatal inactivation of endothelial PP2A also contributes to cognitive dysfunction and neuronal loss. However, regaining endothelial PP2A activity by overexpressing Ppp2ca rescued neuronal dysfunction. Mechanistically, PP2A deficiency is intricately linked to the MPST-H₂S signaling pathway. A robust reduction in endothelial MPST-dependent H₂S production followed PP2A deficiency. Exogenous H₂S treatment and AAV-mediated overexpression of MPST in brain endothelial cells significantly mitigated neuronal dysfunction in PP2A^EC-cKO mice. Furthermore, PP2A deficiency promotes an increase in calcium influx and calpain2 phosphorylation, subsequently leading to MPST degradation. The PP2A activator (FTY720) and MPST activator (3MP sodium) both remarkably restored endothelial MPST-dependent H₂S production, subsequently rescuing ischemia-induced neurological deficits. In conclusion, our study demonstrates that endothelial PP2A deficiency leads to MPST degradation by activating calpain2, thus damaging neuronal function.

ObjectiveThis study aimed to investigate the intervention effect of Renqing Mangjue on the malignant transformation of gastric mucosal epithelial cells induced by N-methyl-N′-nitro-N-nitrosoguanidine （MNNG） and to explore its molecular mechanism in preventing precancerous lesions of gastric cancer based on the cyclic guanosine monophosphate （cGMP）/protein kinase G （PKG）/mitogen-activated protein kinase （MEK）/extracellular signal-regulated kinase （ERK） signaling pathway. MethodsHuman gastric mucosal epithelial cells （GES-1） were initially induced by MNNG to establish a precancerous cell model （MC cells）. The effective concentration of MNNG for inducing malignant transformation in GES-1 cells was screened using the cell proliferation activity decection （CCK-8） assay， and the effective concentration of Renqing Mangjue for inhibiting the proliferation of transformed GES-1 cells was also determined. GES-1 cells were divided into a blank control group， a model group， and treatment groups with Renqing Mangjue at concentrations of 1， 3， 10， and 30 mg·L^-1. Furthermore， the effects of Renqing Mangjue on the migratory ability and epithelial-mesenchymal transition （EMT） characteristics of GES-1 malignant transformed cells were evaluated using Transwell migration assays， wound healing assays， and real-time quantitative reverse transcription polymerase chain reaction （Real-time PCR）. Additionally， candidate chemical components and target sites of Renqing Mangjue were obtained from the TCMIP v2.0 database， and disease targets at various stages of gastric cancer precursors were sourced from the Gene Expression Omnibus （GEO） database. Pathway enrichment analysis was performed using the Metascape database to predict the potential mechanisms of action of Renqing Mangjue. Finally， the protective mechanism of Renqing Mangjue against gastric cancer precursors was validated through Western blot analysis. ResultsAt a concentration of 20 μmol·L^-1， MNNG exhibited an inhibition rate of approximately 50% on GES-1 cells （P<0.01）， and at this concentration， the GES-1 cells displayed biological characteristics indicative of malignant transformation. In contrast， Renqing Mangjue had no significant effect on the proliferation of normal GES-1 cells， but significantly inhibited the proliferation of MC cells （P<0.01） and markedly reduced their migratory capacity （P<0.01）. Moreover， it also increased the mRNA expression level of E-cadherin during the EMT process （P<0.05）， while inhibiting the expression of both N-cadherin and the transcription factor Snail mRNA （P<0.05， P<0.01）. Network predictions suggested that Renqing Mangjue may prevent gastric cancer precursors through modulating the cGMP/PKG and MAPK/ERK signaling pathways. Furthermore， Western blot results indicated that Renqing Mangjue upregulated the expression of PKG and NPRB （B-type natriuretic peptide receptor） proteins in the cGMP/PKG pathway （P<0.01）， while downregulating the expression of the downstream proteins MEK and ERK （P<0.05， P<0.01）. ConclusionIn summary， Renqing Mangjue can prevent gastric cancer precursors by inhibiting the proliferation and migration of malignant transformed GES-1 cells， thereby delaying the EMT process. The underlying mechanisms may be related to the activation of the cGMP/PKG pathway and the inhibition of the MEK/ERK signaling pathway.

Objective:To explore the feasibility and effect on the reconstruction of whole hand degloving injury by transfer of nerved tissue flaps in staged surgery.Methods:A retrospective study was conducted on the clinical data of 5 patients who suffered whole hand degloving injury and underwent staged reconstructive surgery with nerved tissue flaps, from December 2018 to December 2022 in the Department of Hand Surgery, Longgang Orthopaedics Hospital of Shenzhen. The patients were 4 males and 1 female, aged 22-45 years. Two of the whole-hand degloved injuries were left hands and 3 of right. Two patients had the whole-hand degloving injury combined with a fracture of distal phalangeal tuberosity, and 1 was complicated with partial rupture of the extensor tendon insertion. Areas of the whole hand degloving injury ranged from 215 cm 2 to 480 cm 2, the size of the hallux nail flaps for reconstruction of thumbs ranged from 54 cm 2 to 104 cm 2, the size of the hallux nail flaps for reconstruction of index fingers ranged from 65 cm 2 to 133 cm 2, and the size of the flaps for reconstruction of all the defects of hands ranged from 119 cm 2 to 255 cm 2. In primary surgery, the thumbs, index fingers and the first webs were reconstructed with bilateral hallux nail flaps to shape the appearance and gain the sensation function. Meanwhile, a single and large defect was created from the defects of hand by bundling up the middle, ring and little fingers together with the all the defects in both palmar and dorsal hand. Then an anterolateral thigh flap (ALTF) was used to have the created single defect wrapped together. Donor sites of the bilateral hallux nail flap were reconstructed with a lobulated ALTF from the other side or with bilateral peroneal artery perforator flaps. Donor sites of the ALTF and peroneal artery perforator flap were pulled and sutured. After the hallux nail flaps and ALTFs of the affected hands had survived and stabilised, multiple staged surgery were then carried out to firstly reconstruct the ring and little fingers, and followed by the middle and ring fingers in turns from the artificial syndactyly created in the primary surgery. In the final stage of surgery, skin of the radial side of middle and ring finger-pulps and the ulnar little finger-pulp were replaced by lateral toe flaps to reconstruct the sensations of the main sensory zones of middle, ring and little finger-pulps. Thereafter, the shape, TPD and finger extension and flexions were observed and evaluated through the postoperative follow-up, at the outpatient clinic according to the Evaluation Standard of Thumb and Finger Reconstruction Function of the Hand Surgery Society of the Chinese Medical Association. The appearance and function of the donor sites in both feet were evaluated with the Maryland foot score. Results:All flaps survived after surgery. Postoperative follow-up lasted up to 14 to 48 months after the last surgery. The appearance of fingers was satisfactory with good function. TPD of thumbs and index finger-pulps had achieved up to 6-8 mm, and 3-8 mm in the main sensory zones of middle, ring and little finger-pulps. TPD in the non-major sensory zones of middle, ring and little finger-pulps was found at 10-14 mm, which scored 13 to 14 and rated as excellent according to the Evaluation Standard of Thumb and Finger Reconstruction Function of the Hand Surgery Society of the Chinese Medical Association. Only a linear scar left in the donor site of thigh. As the appearance of the flaps on the feet was not bloated and there was no obvious abnormality in walking and running, therefore the function of feet scored up to 96 to 97 and rated excellent according to the Maryland foot score.Conclusion:A multi-staged reconstruction of a whole hand degloving injury with nerved tissue flaps not only achieves satisfactory digital and hand appearance, but also with good function. There is no obvious effect on the appearance and function of the donor sites. This surgical strategy is novel in the reconstruction of a whole hand degloving injury.