Preimplantation polygenic testing， as a frontier technology in assisted reproduction， provides a new way to prevent genetic diseases and reduce birth defects. However， it may also trigger a series of ethical issues. Combined with the research progress of preimplantation polygenic testing， this paper explored the ethical issues that may be caused by this technology， including the risks and uncertain benefits for future offspring and the challenges of informed consent， the dilemmas of autonomous decision-making and genetic responsibility of prospective parents， as well as social risks such as the distortion of the view of life and health inequities. Given the technical safety issues and ethical challenges posed by preimplantation polygenic testing， it was proposed to revise the regulations related to preimplantation genetic testing， as well as improve the ethical review of preimplantation polygenic testing， genetic counseling services， and public education on genetic science and genetic ethics. These ethical countermeasures aim to prevent and mitigate risks and ensure that technology truly benefits human society.

Objective:To establish a prognostic prediction model for hepatocellular carcinoma (HCC) using bioinformatics approaches to guide personalized therapy.Methods:Based on bioinformatics, the differential analysis was carried out on the GSE19665 data set of The Cancer Genome Atlas-Liver Hepatocellular Carcinoma (TCGA-LIHC) and Gene Expression Omnibus (GEO), and the same differentially expressed genes were obtained by means of Wayne diagram. Functional enrichment analyses using Gene Ontology, Kyoto Encyclopedia of Genes and Genome, and Gene Set Enrichment Analysis were conducted on co-expressed genes. Based on clinicopathological and transcriptomic profiles, TCGA-LIHC patients were stratified into training ( n=246) and internal validation ( n=116) cohorts, with external validation using Japanese liver cancer data ( n=231) from the International Cancer Genome Consortium. A LASSO-Cox regression-derived risk scoring model was established and visualized as a nomogram. The clinical utility of the risk score was evaluated through multiple analytical approaches.A nomogram incorporating the risk score was developed, and its predictive performance was validated using the concordance index (C-index) and calibration curves. The measurement data of normal distribution were expressed as mean±standard deviation( ± s), and the t-test was used for comparison between groups. The measurement data with non-normal distribution were expressed as M( Q1, Q3), and the Wilcoxon test was used for comparison between groups. The Kruskal-Wallis test was applied to evaluate the significance of the differences among multiple groups. The prognostic value of the risk score was estimated using Kaplan-Meier analysis and ROC curve. Multivariate Cox regression clarified the independent prognostic value of the risk score. Results:Differential analysis identified 457 commonly expressed differentially expressed genes (DEGs). Enrichment analysis revealed that these common DEGs were significantly enriched in pathways related to the cell cycle of tumor cells.The LASSO-Cox regression model selected eight candidate genes ( CENPA, NDC80, ANXA10, NEIL3, G6 PD, MCM10, SOCS2, MMP1). The predictive risk score generated using these eight genes demonstrated a strong association with the overall survival of HCC patients.The nomogram combining the predictive risk score with clinicopathological features exhibited high predictive performance in both the training and validation cohorts. Furthermore, the prognostic value of this risk score was successfully validated in the external validation cohort. Conclusion:This study successfully developed a new predictive model that accurately predicts the 1-year, 3-year and 5-year survival rates of patients with liver cancer. This can serve as a potential tool to help guide patients in personalized treatment.

Cholera toxin B subunit(CTB)can enhance antigen presentation and promote T cell pro-liferation,B cell differentiation and B cell isotype conversion.Moreover,woodchuck hepatitis virus post transcriptional regulatory element(WPRE)can enhance gene expression efficiency by optimi-zing RNA polyadenylation,denuclearization and/or translation.In order to construct porcine epi-demic diarrhea virus like particles(VLPs)chimerized with CTB and WPRE and evaluate their im-munogenicity,the G Ⅱ type PEDV S gene,combined with the elements promoting the protein ex-pression and enhancing immune effects,was synthesized by the company and cloned into pET32a(+).Af-ter double enzyme digestion and gel recovery,the gene named as TSCW was cloned into pFastBacl to construct the recombinant plasmid pFastBac-TSCW.pFastBac-TSCW was further transformed into DH10Bac competent cells to obtain recombinant bacmid Bacmid-TSCW.Subsequently,the Bacmid-TSCW was transfected into sf9 cells to obtain recombinant baculovirus BV-TSCW.After co-infection of BV-TSCW and BV-M into sf9 cells,viral like particles VLP-TSCW was obtained and used to immunize mice to evaluate its immunogenicity.The results showed that the recombi-nant plasmid pFastBac-TSCW and bacmid Bacmid-TSCW were successfully constructed.After transfection of sf9 cells with recombinant baculovirus,significant cytopathic effects were observed.PCR and Western blot results showed that the recombinant baculoviruses existed stably in sf9 cells and the target proteins was also expressed stably.In addition,the electron microscopy results showed that BV-TSCW and BV-M successfully assembled into viral like particles VLP-TSCW.Furthermore,ELISA results indicated that VLP-TSCW induced high level specific antibodies.The above results laid the foundation for further optimization,design and development of PEDV VLPs subunit vaccines.

The purpose of this study is to investigate the effect of TPCK trypsin on the proliferation pattern of porcine epidemic diarrhea virus in Vero cells.The TPCK trypsin and conventional tryp-sin were added for virus proliferation,and RT-qPCR technology was used to analyze the changes in virus adsorption and invasion in Vero cells.The replication ability of porcine epidemic diarrhea vi-rus in Vero cells was explored through growth curve drawing,IFA identification,and cell activity detection.The results showed that the optimal concentrations of TPCK trypsin and conventional trypsin were 1 mg/L and 6 mg/L,respectively.The virus showed a decreasing trend with the pro-longation of TPCK trypsin and conventional trypsin pretreatment time.Adding different pancreatic enzymes during the virus proliferation process did not promote the virus invasion in Vero cells.Af-ter 4 h of invasion,the virus particles of each group gradually increased.By plotting the growth curve,it was found that the virus content of the TPCK trypsin group reached its highest level at 24 h(lgTCID50=(6.30±0.14)/0.1 mL),followed by a decreasing trend at 36 h,and the fluorescence intensity produced at 24 h was higher than that of conventional trypsin.In summary,TPCK trypsin has a better promoting effect on the proliferation of porcine epidemic diarrhea virus in Vero cells.It provided theoretical basis for further research on the mechanism of TPCK trypsin affecting porcine epidemic diarrhea virus proliferation,and also provided data support for the isola-tion and purification of porcine epidemic diarrhea virus epidemic strains.

The purpose of this study is to investigate the effect of TPCK trypsin on the proliferation pattern of porcine epidemic diarrhea virus in Vero cells.The TPCK trypsin and conventional tryp-sin were added for virus proliferation,and RT-qPCR technology was used to analyze the changes in virus adsorption and invasion in Vero cells.The replication ability of porcine epidemic diarrhea vi-rus in Vero cells was explored through growth curve drawing,IFA identification,and cell activity detection.The results showed that the optimal concentrations of TPCK trypsin and conventional trypsin were 1 mg/L and 6 mg/L,respectively.The virus showed a decreasing trend with the pro-longation of TPCK trypsin and conventional trypsin pretreatment time.Adding different pancreatic enzymes during the virus proliferation process did not promote the virus invasion in Vero cells.Af-ter 4 h of invasion,the virus particles of each group gradually increased.By plotting the growth curve,it was found that the virus content of the TPCK trypsin group reached its highest level at 24 h(lgTCID50=(6.30±0.14)/0.1 mL),followed by a decreasing trend at 36 h,and the fluorescence intensity produced at 24 h was higher than that of conventional trypsin.In summary,TPCK trypsin has a better promoting effect on the proliferation of porcine epidemic diarrhea virus in Vero cells.It provided theoretical basis for further research on the mechanism of TPCK trypsin affecting porcine epidemic diarrhea virus proliferation,and also provided data support for the isola-tion and purification of porcine epidemic diarrhea virus epidemic strains.

Cholera toxin B subunit(CTB)can enhance antigen presentation and promote T cell pro-liferation,B cell differentiation and B cell isotype conversion.Moreover,woodchuck hepatitis virus post transcriptional regulatory element(WPRE)can enhance gene expression efficiency by optimi-zing RNA polyadenylation,denuclearization and/or translation.In order to construct porcine epi-demic diarrhea virus like particles(VLPs)chimerized with CTB and WPRE and evaluate their im-munogenicity,the G Ⅱ type PEDV S gene,combined with the elements promoting the protein ex-pression and enhancing immune effects,was synthesized by the company and cloned into pET32a(+).Af-ter double enzyme digestion and gel recovery,the gene named as TSCW was cloned into pFastBacl to construct the recombinant plasmid pFastBac-TSCW.pFastBac-TSCW was further transformed into DH10Bac competent cells to obtain recombinant bacmid Bacmid-TSCW.Subsequently,the Bacmid-TSCW was transfected into sf9 cells to obtain recombinant baculovirus BV-TSCW.After co-infection of BV-TSCW and BV-M into sf9 cells,viral like particles VLP-TSCW was obtained and used to immunize mice to evaluate its immunogenicity.The results showed that the recombi-nant plasmid pFastBac-TSCW and bacmid Bacmid-TSCW were successfully constructed.After transfection of sf9 cells with recombinant baculovirus,significant cytopathic effects were observed.PCR and Western blot results showed that the recombinant baculoviruses existed stably in sf9 cells and the target proteins was also expressed stably.In addition,the electron microscopy results showed that BV-TSCW and BV-M successfully assembled into viral like particles VLP-TSCW.Furthermore,ELISA results indicated that VLP-TSCW induced high level specific antibodies.The above results laid the foundation for further optimization,design and development of PEDV VLPs subunit vaccines.

By targeting the key gene csgD involved in the biofilm formation of Escherichia coli, we employed molecular docking and molecular dynamics simulation to screen the active components of Chinese medicine with inhibitory effects on the biofilm formation from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). After the anti-biofilm properties of the active components were validated in vitro, data-independent acquisition (DIA) proteomics was employed to further identify the differential proteins involved in interfering with the biofilm formation of Escherichia coli. The mechanisms of inhibition were explored with consideration to the phenotype. Through virtual screening, we identified four candidate active components, including tannic acid, narirutin, salvianolic acid B, and rosmarinic acid. Among them, tannic acid demonstrated significant inhibitory effect on the biofilm formation of E. coli. The analysis of differential proteins, combined with relevant phenotype validation, suggested that tannic acid primarily affected E. coli by intervening in pilus assembly, succinic acid metabolism, and the quorum sensing system. This study provided a lead compound for the development of new drugs against biofilm-associated infections in the future.
Biofilms/drug effects* ; Escherichia coli/metabolism* ; Molecular Docking Simulation ; Drugs, Chinese Herbal/chemistry* ; Tannins/chemistry* ; Cinnamates/metabolism* ; Benzofurans/chemistry* ; Depsides/metabolism* ; Rosmarinic Acid ; Anti-Bacterial Agents/chemistry* ; Escherichia coli Proteins/genetics* ; Medicine, Chinese Traditional

Objective:To investigate the predictive factors for the outcome of patients with complete recanalization after endovascular treatment (EVT) for acute basilar artery occlusion (ABAO).Methods:Patients with ABAO underwent EVT at Jiangsu Provincial People's Hospital and Nanjing Pukou People's Hospital from January 2015 to December 2022 were included retrospectively. Complete recanalization was defined as a modified Thrombolysis in Cerebral Infarction (mTICI) grade 3 after EVT. The main outcome measure was the clinical outcome evaluated by the modified Rankin Scale at 90 days after onset. 0-2 points were defined as good outcome and >2 points were defined as poor outcome. The secondary outcome measure was death within 90 days after onset. Multivariate logistic regression analysis was used to determine the independent predictive factors for poor outcome and mortality. Receiver operating characteristic (ROC) curve was used to analyze the predictive value of the independent predictive factors for poor outcome or mortality. Results:A total of 73 patients with completed recanalization after EVT for ABAO were enrolled, including 55 males (75.3%), aged 67.2±1.58 years, with the median baseline National Institutes of Health Stroke Scale (NIHSS) score 23, median baseline Basilar Artery on Computed Tomography Angiography (BATMAN) score 7, and median baseline posterior circulation Alberta Stroke Program Early CT Score (pc-ASPECTS) 8. After 90 days of onset, 34 patients (46.6%) had poor outcome and 16 (21.9%) died. Multivariate logistic regression analysis showed that higher baseline NIHSS score (odds ratio [ OR] 1.151, 95% confidence interval [ CI] 1.041-1.273; P=0.006), lower baseline pc-ASPECTS ( OR 0.096, 95% CI 0.024-0.386; P=0.001), lower baseline BATMAN score ( OR 0.394, 95% CI 0.162-0.961; P=0.041), and non-first-pass recanalization ( OR 5.011, 95% CI 1.675-23.343; P=0.016) were independently associated with the poor outcome. ROC curve analysis showed that the area under the curve for predicting poor outcome by combining these independent predictive factors was 0.966 (95% CI 0.930-0.964). Multivariate logistic regression analysis showed that older age ( OR 1.147, 95% CI 1.010-1.303; P=0.035), higher baseline NIHSS score ( OR 1.236, 95% CI 1.040-1.470; P=0.016), lower baseline pc-ASPECTS ( OR 0.011, 95% CI 0.002-0.249; P=0.015), and lower baseline BATMAN score ( OR 0.050, 95% CI 0.004-0.618; P=0.020) were independently associated with mortality within 90 days after onset. ROC curve analysis showed that the area under the curve for predicting mortality by combining these independent predictive factors was 0.948 (95% CI 0.899-0.997). Conclusion:For patients with ABAO who had complete recanalization after EVT, the baseline NIHSS score, baseline pc-ASPECTS, baseline BATMAN score, and non-first-pass recanalization are the independent predictive factors for poor outcome at 90 days after onset, while age, baseline NIHSS score, baseline pc-ASPECTS, and baseline BATMAN score are the independent predictive factors for mortality within 90 days after onset.

Accurate and efficient upper limb movement control is a critical guarantee for astronauts to complete their daily tasks in space.Exploring the laws and mechanisms of the influence of microgravity and nonspecific stressors(isolation,noise,fatigue,etc.)in spaceflight on astronauts'upper limb movement control is an important direction of spaceflight human factors engineering research.This article summarizes the research paradigms and findings of in-orbit upper limb movement control research,revealing patterns such as slowed movements and decreased motor control performance under high cognitive load in spaceflight.It also points out the potential mechanisms underlying the inconsistent research results under various research paradigms.On this basis,the paper addresses existing controversies and shortcomings in previous studies,and puts forward prospects and suggestions for subsequent in-orbit movement control research.

Objective:To investigate clinical curative effect of multiple composite tissues replantation with microsurgical technique in single hand.Methods:From May 2008 to December 2021, the clinical data of patients who underwent multiple composite tissue replantation in single hand in Chongming Hospital Affiliated to Shanghai University of Medicine & Health Sciences and Taizhou Hospital of Zhejiang Province, Wenzhou Medical University, were retrospectively analyzed. According to the severed site, anatomical level, the injury cause, the severity of injury, the size of severed tissue, and replantation conditions (such as blood vessels for anastomosis), the severed tissue was evaluated and carried out modified classification (type Ⅰ-Ⅳ) before the operation, and the surgical plan was designed. The severed composite tissue was debrided in emergency, the severed bone and muscle tissue were fixed, and the arteries, veins and nerves were anastomosed by microsurgical technique under a high-power microscope. The ratio of arteries to veins was 1∶1-1∶2. If enough arteries and veins could not be found, arteriovenous shunt could be used to form arterial venous flap or venous arterialized flap, or form artery flap by just artery anastomosis, or form venous flap by just vein anastomosis. Then skin tissue was sutured after recanalization. After the operation, the patients received specialized treatment and guided rehabilitation exercise. The wound healing of hand, skin color, skin quality, swelling degree, sensation, pain degree of joint, and activity function were followed up after the operation. At the last follow-up, the patient’s self-evaluation of the treatment effect was divided into three levels: satisfactory, average, and unsatisfactory. One chief physician and one doctor-in-charge were responsible for the comprehensive evaluation of curative effect, according to the assessment standard for upper limb function issued by Chinese Medical Association of Hand Surgery. 80-100 points were excellent, 60-79 points were good, 40-59 points were fair, and <40 points were poor.Results:A total of 94 severed composite tissues in 35 patients, including of 20 male patients and 15 female patients were enrolled. The average age was 36.4 years (range, 16-65 years). Fifty severed tissues in 18 cases were classified into type Ⅰ, 40 severed tissues in 15 cases type Ⅱ, 4 severed tissues in 2 cases type Ⅲ. Among the 35 cases, 1 case of thumb pulp severed tissue was necrotic at the edge (0.3 cm×0.5 cm) due to local contusion, 1 case of index finger lateral severed tissue was necrotic at the edge (0.3 cm×0.8 cm) due to local avulsion injury, and healed after dressing change, and the other replanted composite tissue survived. The patients were followed up for 3 to 65 months (average, 18.5 months). The wounds were completely healed, and the color, elasticity, and quality of the replanted tissue was close to the surrounding normal skin. There was no obvious swelling or atrophy deformity of the hands. According to the patients’ self-assessment, 31 cases were satisfied and 4 cases were average. Comprehensive evaluation of hand function and appearance: excellent in 23 cases, good in 9 cases, fair in 3 cases, excellent and good rate was about 91.4% (32/35).Conclusion:Though multiple composite tissues replantation with microsurgical technique in single hand is difficult, and vascular anastomosis technical requirement is high, once surgery succeeds, the function and appearance can get good recovery, and patient satisfaction is also relatively high.