ObjectiveA mouse model of vaccinia virus Tiantan strain (VTT)-induced encephalopathy was developed using AG6 mice. MethodsVTT was amplified by infecting Vero cells at a multiplicity of infection (MOI) of 0.01, followed by concentration and titration. After 72 h of incubation, virus-containing cells were collected and subjected to concentration. The concentrated viral suspension was serially diluted (10-fold dilutions) and added to 6-well plates containing confluent Vero cell monolayers for plaque assay. The number of plaques formed in each well was counted, and the virus titer was calculated based on the dilution factor. Fourteen 5-6-week-old AG6 mice (half male and half female, housed separately by sex) were randomly divided into a control group (n=3, PBS), a low-dose group (n=6, 1×10⁵ PFU), and a high-dose group (n=5, 5×10⁵ PFU). The mice were anesthetized by isoflurane inhalation and then infected via intranasal instillation. The mental state of the mice in each group was observed daily, and the body weight and mortality were recorded. On day 13 post-infection, 2% Evans Blue (4 mL/kg body weight) was administered via tail vein injection to assess blood-brain barrier (BBB) disruption. Subsequently, brain tissue samples were collected for immunofluorescence analysis to evaluate the activation of astrocytes and microglia. ResultsThe titer of purified VTT was 1×10⁷ PFU/mL. Compared with the control group, mice in the low-dose group showed no significant change in body weight, and no lethality was observed. In contrast, mice in the high-dose group exhibited significant weight loss starting on day 5 post-infection (P<0.05), accompanied by lethality. On day 13 post-infection, no Evans Blue extravasation was detected in the brain tissues of the low-dose group, while the olfactory bulb region of the high-dose group displayed distinct blue staining, indicating disruption of the BBB. Immunofluorescence analysis revealed no significant proliferation of astrocytes and microglia in the olfactory bulb region of the low-dose group on day 13 post-infection. In contrast, marked activation of glial cells was observable in the high-dose group. ConclusionAn animal model of VTT-induced encephalopathy in AG6 mice is successfully established, characterized by BBB disruption and reactive gliosis specifically localized to the olfactory bulb region, manifested as astrocytic and microglial proliferation.

To investigate the association between the systemic inflammatory response index (SIRI) and lung cancer prevalence based on the National Health and Nutrition Examination Survey (NHANES) database.

Objectives:To analyze the correlation between cervical spine postures and different types of odontoid fractures when injured in different directions of violence using finite element method.Methods:Based on the experience of establishing the finite element model of the upper cervical vertebrae in the early stage,and verified as effective through literature published abroad and in vitro experiments,different positions of the cervical spine(neutral position,flexion position,extension position,lateral bending position,and rotation position)were preset on the established three-dimensional nonlinear finite element model of the normal occip-ital-atlantoaxial complex,and an acceleration of 8m/s2 was applied in each direction of violence[anterior,pos-terior,lateral,top(vertical downward)].The von Mises stress values in different regions of different types(An-derson and D'Alonzo type)of odontoid process fractures were calculated,and stress contour maps were ex-tracted.Results:When subjected to anterior violence,the maximum von Mises stress values were generated in an extension position,with stress concentration in the vertebral body of the axis(Type Ⅲ)(0.41MPa).When subjected to posterior violence,the maximum von Mises stress values were generated when the cervical vertebrae were in a flexion position,with stress concentration in the odontoid process base(Type Ⅱ)(0.52MPa)and the vertebral body of the axis(Type Ⅲ)(0.55MPa).When subjected to lateral violence,the maximum von Mises stress values were generated when the cervical vertebrae were in a flexion position,with stress concen-tration in the odontoid process base(Type Ⅱ)(0.51MPa)and the vertebral body of the axis(Type Ⅲ)(0.43MPa).When subjected to vertical downward violence,the stress values in each body position increased significantly,the stress at the tip of the odontoid process(Type Ⅰ)in the flexion position reached 0.99MPa,the stress at the base of the odontoid process(Type Ⅱ)in the lateral flexion position reached 1.16MPa,and the stress at the axis vertebral body(Type Ⅲ)in the rotational position was 0.85MPa.Conclusions:When the upper cervi-cal spine is in a flexion position and receives a vertically downward force,it is prone to odontoid process tip fractures(Type Ⅰ);When the upper cervical spine is in a flexion position and receives a posterior or lateral force,or when it is in a lateral bending position and receives a vertically downward force,it is prone to odontoid process base fractures(Type Ⅱ);When the upper cervical spine is in a flexion position and receives a posterior force,or when it is in a rotational position and receives a vertically downward force,it is prone to axis vertebral body fractures(Type Ⅲ).

AKT,also known as Protein Kinase B(PKB),plays a critical role in cell proliferation and metabolism.There are three isoforms of AKT:AKT1,AKT2,and AKT3.The effects of these isoforms on the pluripotency and differentiation of mouse embryonic stem cells(mESCs)remain unclear.This study aims to explore the impact of three AKT isoform-specific knockouts on the self-renewal and differen-tiation of mouse embryonic stem cells.Using CRISPR/Cas9 gene-editing technology,AKT isoform-spe-cific knockout cell lines were established.The phenotypic and molecular changes were analyzed through Western blotting,flow cytometry,qRT-PCR,CCK-8 assays,Alkaline Phosphatase(AP)staining,and RNA-seq.The construction of AKT isoform-specific knockout cell lines was successful.The loss of AKT1 and AKT2 inhibited the proliferation of mESCs.The knockout of any single AKT isoform did not affect the expression of pluripotency genes at both mRNA or protein levels.However,during embryoid body forma-tion,the deletion of any of the three AKT isoforms affected the mRNA expression levels of genes in all three germ layers.Transcriptome analysis showed that compared to wild-type mESCs,995,547,and 429 differentially expressed genes(|log2FC|≧1,P＜0.05)were identified inAKT1,AKT2,and AKT3 isoform-specific knockout cells,respectively.There was some overlap in the differentially expressed genes regulated by these three isoforms.In conclusion,the independent knockout of AKT isoforms does not af-fect the maintenance of pluripotency in mouse embryonic stem cells,but they are crucial for differentia-tion.The three AKT isoforms can collectively regulate gene expression while retaining their own regulato-ry specificity.This study provides a foundation for understanding the unique and overlapping roles of AKT isoforms in stem cell biology,highlighting their importance in maintaining stem cell function and differen-tiation.

This study aims to investigate the effects of aerobic exercise on neuroinflammation in AD mice and explore the mechanisms of neuroinflammation regulated by the blood-brain barrier,lipopolysaccharide(LPS)displacement,and glial cell activation.Twenty 3-month-old male APP/PS1 double transgenic mice were used,which were randomly divided into a sedentary group(SE-AD)and an aerobic exercise group(Run-AD),and 10 3-month-old male C57BL/6 mice were used as the control group(WT).The Run-AD group underwent 12 weeks of aerobic training.The results of the water maze showed that aerobic exercise improved the learning and memory capacity of AD mice(P＜0.05).The results of H&E stai-ning and Nissl staining showed that aerobic exercise reduced necrotic cells and inflammatory cell infiltra-tion in the cerebral cortex,as well as nuclear condensation in the CA1 and GD regions of the hippocam-pus(P＜0.05,P＜0.01),and increased the area of Nissl bodies in the cerebral cortex and hippocam-pal CA3 and DG regions.Western blotting and ELISA results showed that aerobic exercise increased the expression of Occludin,ZO-1 and Claudin-5 proteins in the brain(P＜0.01),and decreased the levels of LPS in the brain(P＜0.01).The qRT-PCR results exhibited that aerobic exercise decreased the ex-pression of TLR4,MyD88,NF-κB,IL-1β,and TNF-α mRNA(P＜0.05,P＜0.01).The results of immunofluorescence staining revealed that aerobic exercise reduced the fluorescence area of brain IL-1βand TNF-α proteins(P＜0.05,P＜0.01),as well as the fluorescence area of Iba-1,GFAP,and TLR4 proteins in the cerebral cortex and hippocampus(P＜0.05,P＜0.01).There was a high degree of overlap between Iba-1 and TLR4 fluorescence in the cerebral cortex,and GFAP was localized around Iba-1.In summary,aerobic exercise attenuates neuroinflammation in AD mice by protecting the blood-brain barrier,reducing the displacement of LPS,and subsequently weakening the immune activation of microglia to regulate the TLR4/MyD88/NF-κB signaling pathway to alleviate neuroinflammation.

Pulmonary alveolar proteinosis(PAP)is a rare progressive respiratory dysfunction disease of the lung characterized by insidious onset and non-specific clinical manifestations,often leading to misdiagnosed and mistreated.Herein,we reported a case of PAP patient admitted to Jiading District Central Hospital with an atypical appearance of alveolar lavage fluid and whose condition improved significantly after treatment with subcutaneous injection of recombinant human granulocyte-macrophage colony stimulating factor(GM-CSF).Additionally,we have reviewed and summarized the relevant literature to enhance the understanding of the diagnosis and treatment of this disease.

Pulmonary nodules(PNs),a critical imaging indicator for early lung cancer screening,require deeper mechanistic explo-ration of their malignant transformation.Current Western medicine strategies—primarily surveillance—suffer from delayed interven-tion,while traditional Chinese medicine(TCM)theories(e.g.,"phlegm-blood stasis binding")inadequately explain the dynamic pro-gression of malignancy.Integrating You Yi's theory of"where there is mass hardness,there must be latent yang"with modern research on PN malignant transformation,this study proposes a novel pathogenic mechanism of"latent yang as cause,mass hardness as effect":Depressed latent yang drives pro-tumorigenic microenvironments(e.g.,immunosuppression,hypoxic stress,chronic inflammation),leading to the coagulation of phlegm,stasis,fire,and toxins into cancer toxin.Based on this framework,a core therapeutic principle of"dispersing latent yang and intercepting malignant transition in stages"was established,providing a new paradigm for TCM-based early intervention against PN malignancy.

Pulmonary nodules(PNs),a critical imaging indicator for early lung cancer screening,require deeper mechanistic explo-ration of their malignant transformation.Current Western medicine strategies—primarily surveillance—suffer from delayed interven-tion,while traditional Chinese medicine(TCM)theories(e.g.,"phlegm-blood stasis binding")inadequately explain the dynamic pro-gression of malignancy.Integrating You Yi's theory of"where there is mass hardness,there must be latent yang"with modern research on PN malignant transformation,this study proposes a novel pathogenic mechanism of"latent yang as cause,mass hardness as effect":Depressed latent yang drives pro-tumorigenic microenvironments(e.g.,immunosuppression,hypoxic stress,chronic inflammation),leading to the coagulation of phlegm,stasis,fire,and toxins into cancer toxin.Based on this framework,a core therapeutic principle of"dispersing latent yang and intercepting malignant transition in stages"was established,providing a new paradigm for TCM-based early intervention against PN malignancy.

AKT,also known as Protein Kinase B(PKB),plays a critical role in cell proliferation and metabolism.There are three isoforms of AKT:AKT1,AKT2,and AKT3.The effects of these isoforms on the pluripotency and differentiation of mouse embryonic stem cells(mESCs)remain unclear.This study aims to explore the impact of three AKT isoform-specific knockouts on the self-renewal and differen-tiation of mouse embryonic stem cells.Using CRISPR/Cas9 gene-editing technology,AKT isoform-spe-cific knockout cell lines were established.The phenotypic and molecular changes were analyzed through Western blotting,flow cytometry,qRT-PCR,CCK-8 assays,Alkaline Phosphatase(AP)staining,and RNA-seq.The construction of AKT isoform-specific knockout cell lines was successful.The loss of AKT1 and AKT2 inhibited the proliferation of mESCs.The knockout of any single AKT isoform did not affect the expression of pluripotency genes at both mRNA or protein levels.However,during embryoid body forma-tion,the deletion of any of the three AKT isoforms affected the mRNA expression levels of genes in all three germ layers.Transcriptome analysis showed that compared to wild-type mESCs,995,547,and 429 differentially expressed genes(|log2FC|≧1,P＜0.05)were identified inAKT1,AKT2,and AKT3 isoform-specific knockout cells,respectively.There was some overlap in the differentially expressed genes regulated by these three isoforms.In conclusion,the independent knockout of AKT isoforms does not af-fect the maintenance of pluripotency in mouse embryonic stem cells,but they are crucial for differentia-tion.The three AKT isoforms can collectively regulate gene expression while retaining their own regulato-ry specificity.This study provides a foundation for understanding the unique and overlapping roles of AKT isoforms in stem cell biology,highlighting their importance in maintaining stem cell function and differen-tiation.

Objective To study the predictive value of umbilical artery blood gas analysis combined with early postnatal myocardial enzyme spectrum for neonatal respiratory distress syndrome(RDS)and its severity in premature infants.Methods From January 2018 to December 2024,85 newborns with the gestation age of 30-36+6 weeks who were diagnosed as RDS after birth form Hangzhou Women's Hospital were selected as study group.85 newborns without diagnosed as RDS during the same period were included as control group in a 1:1 ratio.All of them born by cesarean section.The umbilical arterial blood gas analysis was performed and blood samples were collected for blood biochemical tests.The umbilical artery blood pH,base excess,lactate,and myocardial enzyme spectrum such as aspartate aminotransferase(AST),lactate dehydrogenase(LDH),creatine kinase isoenzymes(CK-MB)were compared between two groups.Binary Logistic regression was used to analyze the correlation between umbilical artery blood gas analysis and venous blood myocardial enzyme spectrum and RDS.Receiver operating characteristic curve of umbilical artery blood pH value and creatine kinase isoenzyme alone and their combined detection for predicting RDS was detected,and area under the curve(AUC),sensitivity and specificity of the single detection and combined detection were compared.The study group was further divided into subgroup A(mild)and subgroup B(severe)and the differences of the above indicators between two subgroups were compared.Results Compared with control group,tstudy group had lower pH and base excess levels in umbilical artery blood as well as AST,LDH and CK-MB were higher than those in control group with statistically significant differences(P＜0.05).pH and CK-MB were correlated with RDS(P＜0.05).The AUC of umbilical artery blood pH in predicting RDS was 0.728,the sensitivity was 0.588 and the specificity was 0.812.Combined with CK-MB,the AUC of predicting RDS was 0.813,the sensitivity was 0.729,and the specificity was 0.824.In the study group,subgroup A had higher pH and base excess levels in umbilical artery blood as well as AST,LDH and CK-MB were lower than those in subgroup B with statistically significant differences(P＜0.05).CK-MB was correlated with disease severity in newborns with RDS(P＜0.01).Conclusion Compared with simple application of pH,the combined use of CK-MB can more accurately predict RDS in premature infants,and also has a certain significance for the severity of RDS in premature infants.