Background: and Purpose Therapeutic target selection in noninvasive brain stimulation for poststroke motor recovery typically relies on the interhemispheric inhibition model, which is effective for mildly affected patients but offers limited benefits for severely affected individuals. The mechanisms governing recovery from moderate-to-severe stroke remain poorly understood, which hinders the development of targeted interventions. Methods: We analyzed resting-state functional magnetic resonance imaging data from patients with unilateral subcortical stroke and moderate-to-severe upper limb deficits, both pre- and postintervention, along with data from healthy controls. We developed a novel dynamic lag analysis method for identifying recovery-related homotopic sensorimotor regions with altered interhemispheric interactions. To further uncover the global reorganization pathway, we developed dynamic lateralization approaches to detect large-scale functional connectivity (FC) alterations associated with the identified regions in transient lateralization states. Results: Dynamic time-lag analysis revealed significantly reduced synchronized states in the homotopic dorsal premotor cortex (PMd) post-intervention compared with pre-intervention, which correlated with motor recovery. Further dynamic lateralization analysis revealed a prolonged segregation state in patients, characterized by weakened interhemispheric and strengthened intrahemispheric interactions. In this state, patients showed decreased FC in the ipsilesional PMd and increased FC in the contralesional PMd with bilateral subcortical networks. These recoveryrelated alterations were absent in the traditional static analysis. Conclusions Dynamic analyses targeting interhemispheric interactions are valuable for understanding neural reorganization after stroke. The diminished interactions between the homotopic PMd indicate a compensatory mechanism. Importantly, a state-dependent compensatory pathway was identified, wherein the contralesional PMd assumes the functions of the ipsilesional PMd through enhanced interactions with subcortical structures, potentially guiding more effective interventions.

Genetically engineered mice are important tools for elucidating gene function and disease mechanisms and for advancing drug development. With the growing demand in biomedical research for complex genetic modifications, rapid model generation, and high-throughput functional analysis, technologies for generating genetically engineered mice continue to evolve. This review systematically summarizes the evolution and applications of these technologies, with a focus on conventional approaches, including early transgenic technologies and embryonic stem cell (ESC)-based homologous recombination for gene targeting, as well as the use of random mutagenesis approaches, such as N-ethyl-N-nitrosourea (ENU) mutagenesis and transposon insertion, in forward genetic screening. It further discusses the principles and characteristics of programmable nucleases, including zinc finger nucleases, transcription activator-like effector nucleases, and CRISPR/Cas systems, as well as their applications in generating genetically engineered mice. With advances in genome editing, Cas9-derived systems, base editing, prime editing, and RNA editing further improve editing precision, expand the repertoire of genetic modifications and precise regulatory approaches, and provide additional options for generating genetically engineered mice. This review also compares the advantages, limitations, and suitability of different technologies for specific applications and summarizes quality-control strategies addressing practical issues that may affect model reliability, including off-target effects, mosaicism, genetic background, and genetic stability. Finally, future directions in technologies for generating genetically engineered mice are discussed to inform the generation and application of related models.

Objective: To explore the mechanism of action of hepatic leukemia factor (HLF) in diabetes mellitus and to construct a conditional animal model of mice with islet β ⁃cell⁃specific HLF gene knockout. Methods: At the cellular level , the effects of HLF inhibition or overexpression on the proliferation of MIN6 cells was verified by the CCK⁃8 assay. The effects of HLF inhibition or overexpression were detected at the mRNA level and protein level by Cre + / - mice (C57BL/6J) to obtain offspring mice. The genotypes of the mice were identified by the PCR method.The differences in the expression levels of the HLF gene at the mRNA and protein levels in islet β ⁃cell knockout mice (HLFflox/flox Cre + / - ) and control mice (HLFflox/flox ) were detected by RT⁃qPCR technology and Western blot technology to verify the knockout effect. At the same time , the islet tissues of the mice in two groups were taken to make paraffin sections and analyzed by hematoxylin⁃eosin (HE) staining. Results: HLF gene inhibition or overex⁃pression had no significant effect on the proliferation of MIN6 cells. When the HLF gene was inhibited in MIN6 cells , the mRNA expression level decreased by 74% compared with the control group , and the protein expression level decreased by 60% compared with the control group. After overexpressing the HLF gene , the mRNA expres⁃sion level was 2. 13 times compared with that of the control group , and the protein expression level was 1. 8 times compared with that of the control group. The mRNA expression level of the HLF gene in the knockout mice de⁃creased by 89% compared with the control group , and the protein expression level decreased by 65% compared with the control group. The results of HE staining showed that there was no significant difference in the cell mor⁃phology in the islet tissues between the knockout mice and the control mice. Inhibiting HLF increased the glycogen content in MIN6 cells by approximately 20% . Conclusion The HLF gene knockout mice are successfully con⁃structed , providing an animal model for studying the role of HLF in the pathogenesis of diabetes mellitus.

Objective: To compare the changes in the concentration of relevant growth factors released from platelet-rich plasma (PRP) stored at -80℃ by cryopreservation and at 4℃ by refrigerated lyophilization over 2 years, aiming to provide a theoretical basis for prolonging PRP storage duration. Methods: PRP (n=15) was separated using a blood cell separator and stored under -80℃ cryopreservation (F-PRP group) and 4℃ refrigerated freeze-drying conditions (FD-PRP group). The contents of growth factors (PDGF-AA, PDGF-BB, EGF, TGF-β1, and VEGF) in both groups were measured by ELISA at 1, 3, 6, 9, 12 and 24 months. Results: PDGF-AA and VEGF maintained good stability in both groups for up to 24 months. PDGF-BB and TGF-β1 showed high stability in the first 12 months but their stability decreased gradually from 12th to 24th months. EGF demonstrated good stability in the first 6 months, and its stability gradually decreased from the 9th to 24th months. Comparing the F-PRP and FD-PRP groups, the concentrations of the five growth factors in the FD-PRP group were either not statistically different or higher than those in the F-PRP group at all time points. Specifically, the concentrations of EGF were significantly higher in the FD-PRP group at all time points. Conclusion: Both -80℃ freezing and 4℃ freeze-drying enable long-term preservation of PRP. Freeze-drying imposes less stringent storage requirements and facilitates growth factor compared to frozen storage.

Objective: To evaluate the immunogenicity and safety of co-administration with bivalent human papillomavirus (HPV) vaccine and hepatitis E virus (HEV) vaccine, so as to provide reference for optimizing the vaccination schedule. Methods: Females aged 18 to 25 years were recruited from September to October 2021 in Hengdian College of Film & Television in Zhejiang Province and randomly divided into the HPV+HEV group, the HPV group, and the HEV group. The vaccination procedures were one dose each at 0, 1, and 6 months. Immunogenicity was evaluated by detecting the geometric mean titers (GMT) of HPV16 IgG, HPV18 IgG, and/or HEV IgG antibodies before the first vaccination and one month after the full course of immunization, and comparing the difference in seroconversion, and the GMT ratio. The non-inferiority margin was set at a seroconversion difference of ≤5%, and the lower limit of the 95%CI of the GMT ratio was >0.5. Safety was evaluated by collecting conjunctive local reactions/events and systemic reactions/events within 7 days after each dose, non-conjunctive adverse events within 30 days after each dose, and serious adverse events throughout the observation period (0 to 7 months). Results: A total of 240 females were included, among whom 236 completed the full vaccination program, including 79 in the HPV+HEV group, 77 in the HPV group, and 80 in the HEV group. One month after the full course of immunization, the seroconversion rates of HPV16 IgG and HPV18 IgG antibodies in both the HPV+HEV group and the HPV group were 100%, and the differences in seroconversion rates were 0 (95%CI: -3.39%-+∞). The seroconversion rates of HEV IgG antibodies in both the HPV+HEV group and the HEV group were 100%, and the difference in seroconversion rates was 0 (95%CI: -3.27%-+∞). The GMT of HPV16 IgG and HPV18 IgG antibodies in the HPV+HEV group was 393.88 and 284.86 IU/mL respectively, which was not inferior to 489.39 and 341.24 IU/mL in the HPV group, and the GMT ratios were 0.80 (95%CI: 0.66-+∞) and 0.83 (95%CI: 0.68-+∞), respectively. The GMT of HEV IgG in the HPV+HEV group was 13.55 U/mL, which was not inferior to 12.72 U/mL in the HEV group, and the GMT ratio was 1.07 (95%CI: 0.92-+∞). The incidences of pain, pruritus, and induration in the HPV+HEV group were 54.43%, 21.52% and 40.51% respectively, which were significantly higher than 10.39%, 0, and 0 in the HPV group (all P<0.05). The incidences of redness/swelling, muscle pain/general weakness in the HPV+HEV group were 2.53% and 0, respectively, which were significantly lower than 12.50% and 16.25% in the HEV group (both P<0.05). Conclusion The co-administration of the bivalent HPV vaccine and HEV vaccine is not inferior to individual vaccination in terms of immunogenicity and safety, and the vaccination plan can be optimized through co-administration.

Objective To explore the current status and influencing factors of heart failure occurrence in patients with unstable angina pectoris (UAP), and to provide a scientific basis for developing individualized prevention and treatment strategies. Methods A total of 310 patients with UAP admitted to the Fifth People's Hospital from October 2021 to October 2024 were selected as study subjects. The current status of the patients' heart failure was statistically analyzed, and the patients were divided into heart failure group and non-heart failure group according to whether they had heart failure. Univariate and logistic multivariate regression analyses were used to analyze the risk factors for the occurrence of heart failure in patients with UAP. Results Among the 310 patients with UAP, 63 cases had heart failure, with an incidence rate of 20.32%. After logistic multivariate analysis, it was found that diabetes mellitus, hyperlipidemia, number of coronary artery lesions, homocysteine and plasma brain natriuretic peptide levels were risk factors of heart failure in patients with UAP, and hemoglobin level was a protective factor (OR: 2.010, 95%CI: 1.063-3.800; OR: 4.495, 95%CI: 2.228-9.067; OR: 2.408, 95%CI: 1.256-4.617; OR: 3.655, 95%CI: 1.812-7.372; OR: 4.693, 95%CI: 2.622-8.399; OR: 0.359, 95%CI: 0.205-0.628, P<0.05). Conclusion The coronary heart disease risk of heart failure is high in patients with UAP, and is affected by comorbidities, number of coronary artery lesions, homocysteine, and plasma brain natriuretic peptide levels. It is necessary to perform clinical screening and pay attention to such patients, and take active prevention and control interventions.

BACKGROUND: Neuroticism has been associated with numerous health outcomes. However, most research has focused on a single specific disorder and has produced controversial results, particularly regarding mortality risk. Here, we aimed to examine the association of neuroticism with morbidity and mortality and to elucidate how neuroticism affects trajectories from a healthy state, to one or more neuroticism-related disorders, and subsequent mortality risk. METHODS: We included 483,916 participants from the UK Biobank at baseline (2006-2010). Neuroticism was measured using the Eysenck Personality Questionnaire. Three clusters were constructed, including worry, depressed affect, and sensitivity to environmental stress and adversity (SESA). Cox proportional hazards regression and multistate models were used. Linear regression was used to examine the association between neuroticism and immune parameters and neuroimaging measures. RESULTS: High neuroticism was associated with 37 non-overlapping diseases, including increased risk of infectious, cardiometabolic, neuropsychiatric, digestive, and respiratory diseases, and decreased risk of cancer. After adjustment for sociodemographic variables, physical measures, healthy behaviors, and baseline diagnoses, moderate-to-high neuroticism was associated with a decreased risk of all-cause mortality. In multistate models, high neuroticism was associated with an increased risk of transitions from a healthy state to a first neuroticism-related disease (hazard ratio [HR] [95% confidence interval (CI)] = 1.09 [1.05-1.13], P  <0.001) and subsequent transitions to multimorbidity (1.08 [1.02-1.14], P  = 0.005), but was associated with a decreased risk of transitions from multimorbidity to death (0.90 [0.84-0.97], P for trend = 0.006). The leading neuroticism cluster showing a detrimental role in the health-illness transition was depressed affect, which correlated with higher amygdala volume and lower insula volume. The protective effect of neuroticism against mortality was mainly contributed by the SESA cluster, which, unlike the other two clusters, did not affect the balance between innate and adaptive immunity. CONCLUSION This study provides new insights into the differential role of neuroticism in health outcomes and into new perspectives for establishing mortality prevention programs for patients with multimorbidity.
Humans ; Neuroticism/physiology* ; Male ; Female ; Middle Aged ; Aged ; Proportional Hazards Models ; Surveys and Questionnaires ; Adult ; Risk Factors

Medical fiber endoscope is one kind of medical device that utilizes fiber technique to realize visual detection for internal organs or cavities.In clinical applications,medical fiber endoscope not only need to complete fiber optic imaging but also need possess the functions of delivering water and air,and operating device.Therefore,its routine management and maintenance for fault is more and more payed attention by the staffs of using medical fiber endoscope in department.The system of delivering water and air is a key structure for some of medical fiber endoscopes,which has higher maintenance matters and frequency of maintaining fault.Based on the above reasons,this research analyzed the working principle of medical fiber endoscopes,and summarized the ideas and methods of repairing common faults through focused on the structural composition of delivering system of water and air of medical fiber endoscopes,so as to improve the management level for this type of medical device.

Objective To investigate the correlation between the expression of human leukocyte antigen class I(HLA-I)and programmed cell death ligand 1(PD-L1)with clinicopathological features and cellular immune infiltration.Methods A total of 150 patients with bladder cancer diagnosed and treated in Anhui Provincial Hospital from May 2020 to April 2023 were retrospectively selected as the study objects.The positive expression rates and positive scores of HLA-I and PD-L1 were compared between cancerous tissues and adjacent tissues.The positive scores of HLA-I and PD-L1 in cancer tissues of patients with different clinical characteristics were compared,and the correlation between HLA-I,PD-L1 and clinical characteristics of patients with bladder cancer was analyzed by Kendall's tau-b method.Logistic regression model was used to establish the combined model parameters of HLA-I and PD-L1 positive scores,and receiver operating characteristic(ROC)curve was drawn to analyze the HLA-I and PD-L1 positive scores and the area under the curve(AUC),sensitivity and specificity of the combined diagnosis of bladder cancer.Results The positive expression rate of HLA-I in cancer tissues was lower than that in paracancer tissues[38.67%(58/150)vs 81.33%(122/150)],while the positive expression rate of PD-L1 was higher than that in paracancer tissues[57.33%(86/150)vs 14.00%(21/150)],and the differences were statistically siginficant(χ2=56.889,61.377,all P<0.05).The HLA-I positive score of cancer tissues was lower than that of paracancer tissues[2.00(1.00,3.00)vs 3.00(3.00,5.00)],while the PD-L1 positive score was higher than that of paracancer tissues[3.00(2.00,5.00)vs 2.00(1.00,2.00)],and the fifferences were statistically significant(Z=-8.409,-6.346,all P<0.05).There was no significant difference in HLA-I and PD-L1 positive scores among different sex,age and tumor diameter(ZHLA-1=-1.834,-0.622,-0.543;ZPD-L1=0.811,0.812,0.919,all P＞0.05).The difference of HLA-I and PD-L1 positive scores among different pathological stages,lymph node metastasis,differentiation degree,CD4+,CD8+and CD68+were statistically significant(ZHLA-1=-7.034～3.814;ZPD-L1=-4.479～3.257,all P＜0.05).Kendall's tau-b correlation analysis showed that HLA-I was negatively correlated with pathological stage,lymph node metastasis,degree of differentiation,and positively correlated with negative infiltration of CD4+,CD8+and CD68+(r=-0.528～-0.286,all P<0.05).PD-L1 was positively correlated with pathological stage,lymph node metastasis,degree of differentiation and negatively correlated with negative infiltration of CD4+,CD8+and CD68+(r=-0.243～0.334,all P<0.05).ROC curve analysis showed that the positive scores of HLA-I and PD-L1 and the AUC values of the combined diagnosis of bladder cancer were 0.773,0.702 and 0.856,respectively.Sensitivity was 61.30%,57.30%and 82.00%.The specificity was 81.30%,86.00%and 73.30%.Conclusion The expression of HLA-I and PD-L1 is abnormal in patients with bladder cancer,and their expression is affected by the positive infiltration of immune cells.Observing the positive expression of HLA-I and PD-L1 is beneficial to provide guidance for clinical diagnosis and treatment.

Objective:To evaluate the clinical efficacy of botulinum toxin type A (BTX-A) injection in enhancing chin aesthetics.Methods:A retrospective analysis was conducted on patients with suboptimal chin aesthetics who underwent dual-plane BTX-A injection at Plastic Surgery Hospital, Chinese Academy of Medical Sciences between August 2023 and March 2024. Prior to injection, patients were instructed to repeatedly pucker their lips forward and upward to identify the most prominent points of the mentalis muscle for injection. A 13 mm 30 G needle was inserted perpendicularly into the muscle layer, and BTX-A was administered at a concentration of 20 U/ml, with 3 U per injection point. For the patients exhibiting significant orange peel signs at rest, intradermal deep-layer BTX-A injection was concurrently performed at a concentration of 5 U/ml, with 0.5 U per linear track. Postoperative follow-up was conducted, and third-party physicians assessed pre- and post-treatment photographs using the global aesthetic improvement scale (GAIS) (scores ranging from 5 to 1, representing worse, no improvement, mild improvement, moderate improvement, and significant improvement, respectively). Patient satisfaction was also surveyed [categorized as very satisfied, satisfied, dissatisfied, or very dissatisfied; satisfaction rate = (very satisfied + satisfied) cases/total cases × 100%], along with their willingness to undergo repeated injections and recommend the procedure to others. Descriptive statistical analysis was performed using SPSS 24.0 software. Normally distributed continuous data were expressed as Mean±SD, and categorical data were expressed as counts (%).Results:A total of 120 patients were included, comprising 11 males and 109 females, aged 22-39 years (mean age of 33.3 years). Follow-up ranged from 1 to 5 months (mean of 1.3 months). Postoperatively, 102 patients reported subjective improvement in chin appearance, characterized by enhanced fullness and roundness of the chin. Thirty-one patients noted a slight elevation of the submental fat pad and improved definition of the cervicomental angle. The mean GAIS score was 1.61±0.78, with 76 cases scoring 1, 24 cases scoring 2, 10 cases scoring 3, and 10 cases scoring 4. Improvement (scores 1-3) was achieved in 91.7% (110/120) of patients. The subjective satisfaction rate was 85.0% (102/120), and 94 patients (78.3%) expressed willingness to undergo repeated injections and recommend the procedure to family or friends. Early postoperative complications included localized bruising in 17 cases, which was resolved within 10 d, and transient fine motor dysfunction of the lower jaw in 23 cases, with normal chewing, swallowing, and facial expressions, all of which were resolved completely within 6 weeks. No cases of mouth deviation, facial paralysis, allergic reactions, or other complications were observed.Conclusion:The application of BTX-A via intramuscular mentalis injection combined with intradermal deep-layer injection significantly improves both dynamic and static chin aesthetics. However, some common complications also ask for the further attention of practitioners.