ObjectiveTo investigate the mechanisms of Tongnao decoction （TND） in mice with acute ischemic stroke （AIS）. MethodsFifty male C57BL/6J mice were randomly divided into a sham operation group， model group， TND low-dose group （1.86 g·kg^-1）， TND high-dose group （3.72 g·kg^-1）， and butylphthalide （NBP） group （10 mg·kg^-1）， with 10 mice in each group. A mouse model of cerebral ischemic injury was established using photochemical thrombosis （PT）. The sham operation group and model group were administered an equal volume of normal saline by gavage. All five groups were treated once daily for 14 consecutive days. Behavioral tests were performed before modeling and at the end of administration. T₂-weighted imaging （T₂WI） was performed 3 days after modeling to evaluate the extent of injury. Hematoxylin-eosin （HE） staining was used to observe histological changes in the cerebral cortex， and Nissl staining was used to observe neuronal morphology. Cerebral blood flow in mice was detected using a laser speckle contrast imaging （LSCI） system. Immunofluorescence staining was used to detect the cell proliferation marker bromodeoxyuridine （BrdU） and the highly glycosylated type I transmembrane glycoprotein CD34. Western blot analysis was used to detect the expression levels of phosphatidylinositol 3-kinase （PI3K）， protein kinase B （Akt）， glycogen synthase kinase-3β （GSK-3β）， and their phosphorylation levels， as well as tight junction-related proteins zonula occludens-1 （ZO-1）， Occludin， and Claudin-5 in the peri-infarct tissue. Thirty-five zebrafish were randomly divided into normal control group， model group， TND low and high dose groups （0.16， 0.32 g·L^-1） and NBP group （10 μmol·L^-1）， with 7 in each group. A stereoscopic fluorescence microscope was used to observe vascular growth in zebrafish. ResultsImaging showed that PT caused ischemia in the right cortical region. Behavioral tests indicated that， compared with the model group， the drug-treated groups reduced the error rate of irregular balance ladder climbing on the affected side and shortened the tape removal time （P<0.05）. HE staining and Nissl staining showed that， compared with the model group， the drug-treated groups exhibited reduced brain tissue damage， fewer scars， and improved neuronal morphology. LSCI results showed that the drug-treated groups partially restored cerebral blood perfusion and promoted the establishment of collateral circulation compared with the model group. Immunofluorescence staining indicated that the drug-treated groups increased the positive rates of BrdU and CD34 compared with the model group （P<0.01）， promoting angiogenesis. Meanwhile， compared with the model group， the drug-treated groups upregulated the expression levels of p-PI3K， p-Akt， p-GSK-3β， and tight junction proteins ZO-1， Occludin， and Claudin-5 （P<0.05，P<0.01）， and increased the number of intersegmental vessels in zebrafish （P<0.05，P<0.01）. ConclusionTND can promote angiogenesis around the infarct in PT model mice by regulating the PI3K/Akt/GSK-3β signaling pathway， thereby improving cerebral ischemic injury.

The translocator protein (TSPO) positron emission tomography (PET) can noninvasively detect neuroinflammation associated with epileptogenesis and epilepsy. This study explored the role of the TSPO-targeting radioligand [¹⁸F]F-TFQC, an m-trifluoromethyl ER176 analog, in the PET neuroimaging of epileptic rats. Initially, [¹⁸F]F-TFQC was synthesized with a radiochemical yield of 8%-10% (EOS), a radiochemical purity of over 99%, and a specific activity of 38.21 ± 1.73 MBq/nmol (EOS). After determining that [¹⁸F]F-TFQC exhibited good biochemical properties, [¹⁸F]F-TFQC PET neuroimaging was performed in epileptic rats at multiple time points in various stages of disease progression. PET imaging showed specific [¹⁸F]F-TFQC uptake in the right hippocampus (KA-injected site, i.e., epileptogenic zone), which was most pronounced at 1 week (T/NT 1.63 ± 0.21) and 1 month (T/NT 1.66 ± 0.20). The PET results were further validated using autoradiography and pathological analysis. Thus, [¹⁸F]F-TFQC can reflect the TSPO levels and localize the epileptogenic zone, thereby offering the potential for monitoring neuroinflammation and guiding anti-inflammatory treatment in patients with epilepsy.

Purpose To investigate the value of radiomics nomogram for the prediction of p53 abnormal in patient with endometrial carcinoma based on intratumoral and peritumoral MRI.Materials and Methods A total of 145 female patients were pathologically confirmed endometrial carcinoma who underwent pelvic MRI before treatment in Baoding First Central Hospital from January 2020 to April 2024,including 96 patients with p53 wild type and 49 with p53 abnormal.Radiomics features were extracted from both intratumoral and peritumoral regions(2 mm)in diffusion weighted imaging and equilibrium phase of dynamic contrast enhanced MRI,which were selected using least absolute shrinkage and selection operator.Three machine learning algorithms of random forest,K-nearest neighbors and extra trees were conducted to develop the intratumoral,peritumoral and intratumoral combined peritumoral radiomics models.Multivariate Logistic regression was used to constitute the clinical model and nomogram.The performance of these models was evaluated using receiver operating characteristic curve,decision curve analysis and calibration curve.Results The K-nearest neighbors model of the intratumoral combined peritumoral regions performed the best in all radiomics models,the area under the curve were 0.921 and 0.773 in the training cohorts and test cohorts.The radiomics nomogram,which was composed of age,apparent diffusion coefficient and radiomics signatures,achieved the best performance with area under the curve of 0.970 and 0.777 in the training cohorts and test cohorts,respectively.Calibration curve analysis and decision curve analysis demonstrated favorable calibration and clinical utility of the nomogram model.Conclusion The nomogram based on intratumoral and peritumoral MRI radiomics yields a favorable diagnostic value for predicting p53 abnormal in patient with endometrial carcinoma.

Objective:To investigate the protective effects of p53/glucose transporter 4 (GLUT4) regulation on cardiomyocyte injury induced by high glucose combined with hypoxia/reoxygenation.Methods:Human myocardial AC16 cells were treated with 33 mmol/L glucose and a hypoxic chamber to establish an in vitro model of high glucose combined with hypoxia/reoxygenation. Based on the glucose concentration in the medium and hypoxia/reoxygenation conditions, AC16 cells were divided into control group, high glucose group, hypoxia/reoxygenation group and high glucose combined with hypoxia/reoxygenation group. On the basis of high glucose combined with hypoxia/reoxygenation group, cells were transfected with empty vector, p53 small interfering RNA (siRNA), and co-transfected with p53 and GLUT4 siRNA to establish negative control group, sip53 transfection group, and sip53+siGLUT4 transfection group, respectively. Western blotting was used to detect the levels of hypoxia-inducible factor-1α (HIF-1α), p53, GLUT4, dynamin-related protein 1 (Drp1), mitofusin 2 (Mfn2), B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax) and cysteine aspartic acid specific protease-3 (Caspase-3). The levels of reactive oxygen species were detected using the 2′,7′-dichlorodihydrofluorescein diacetate fluorescent probe. Mitochondria were labeled with the Mito-Tracker Deep Red FM fluorescent probe to assess mitochondrial morphology and their related parameters. Mitochondrial membrance potential was meausred using the JC-1 detection kit. Adenosine triphosphate (ATP) content was determined using an ATP assay kit. Glucose uptake ability was evaluated by measuring the fluorescence intensity of 2-[ N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxy- D-glucose (2-NBDG) using a multifunctional microplate reader. Apoptosis was assessed by TUNEL assay. Results:The relative expression of HIF-1α protein in the high glucose combined with hypoxia/reoxygenation group was 1.189±0.185, higher than that in the control group (0.086±0.071) ( P<0.05). The relative expression of p53 protein in the high glucose combined with hypoxia/reoxygenation group was 1.248±0.194, higher than those in the control group (0.730±0.184), high glucose group (0.932±0.161) and hypoxia/reoxygenation group (1.109±0.151) (all P<0.05). The relative expression of GLUT4 protein in the high glucose combined with hypoxia/reoxygenation group was 0.407±0.140, lower than those in the control group (1.061±0.060) and hypoxia/reoxygenation group (0.781±0.092) (both P<0.05). The fluorescence intensity of reactive oxygen species in the high glucose combined with hypoxia/reoxygenation group was 38.31±1.66, higher than that in the control group (11.59±1.02) ( P<0.05). The number of mitochondria in the high glucose combined with hypoxia/reoxygenation group was (62.00±15.26), lower than those in the control group (136.20±23.55) and high glucose group (96.55±13.72) (both P<0.05). The average mitochondrial area in the high glucose combined with hypoxia/reoxygenation group was (7.02±1.38) μm 2, lower than those in the control group [(13.74±0.67) μm 2], high glucose group [(9.27±1.99) μm 2] and hypoxia/reoxygenation group [(9.64±2.36) μm 2] (all P<0.05). The average perimeter of mitochondria in the high glucose combined with hypoxia/reoxygenation group was (9.10±1.14) μm, lower than those in the control group [(13.35±0.69) μm] and the hypoxia/reoxygenation group [(10.83±1.58) μm] (all P<0.05). The number of mitochondrial branches was 53.73±9.49, lower than those in the control group (147.10±25.99), high glucose group (97.08±13.65) and hypoxia/reoxygenation group (104.80±24.92) (all P<0.05). The average branch length of mitochondria in the high glucose combined with hypoxia/reoxygenation group was (1.45±0.26) μm, lower than that in the control group [(2.29±0.52) μm] ( P<0.05). The red-green fluorescence intensity ratio in the high glucose combined with hypoxia/reoxygenation group was 0.580±0.133, lower than those in the control group (2.379±0.242), high glucose group (1.200±0.112) and hypoxia/reoxygenation group (0.883±0.076) (all P<0.05). The ATP content of the high glucose combined with hypoxia/ reoxygenation group was (0.025±0.003) μmol/10 5 cells, lower than those of the control group [(0.137±0.012) μmol/10 5 cells], high glucose group [(0.078±0.003) μmol/10 5 cells] and hypoxia/reoxygenation group [(0.073±0.010) μmol/10 5 cells] (all P<0.05). The fluorescence intensity of 2-NBDG in the high glucose combined with hypoxia/reoxygenation group was 257 315±7 951, lower than those in the control group (339 597±10 165), high glucose group (317 293±8 876) and hypoxia/reoxygenation group (314 611±12 228) (all P<0.05). The relative expression of Drp1 protein in high glucose combined with hypoxia/reoxygenation group was 1.203±0.090, higher than those in the control group (0.705±0.170), high glucose group (0.910±0.106) and hypoxia/reoxygenation group (1.002±0.112) (all P<0.05). The relative expression of Mfn2 protein in the high glucose combined with hypoxia/reoxygenation group was 0.706±0.285, lower than those in the control group (1.988±0.139), high glucose group (1.305±0.076) and hypoxia/reoxygenation group (1.131±0.236) (all P<0.05). The relative expression levels of Bax/Bcl-2 and Caspase-3 proteins in the high glucose combined with hypoxia/reoxygenation group were 2.318±0.216 and 1.076±0.076, respectively, higher than those in the control group (0.281±0.046 and 0.442±0.084), high glucose group (0.673±0.043 and 0.662±0.159) and hypoxia/reoxygenation group (0.807±0.293 and 0.835±0.058), respectively (all P<0.05). The TUNEL fluorescence intensity of the high glucose combined with hypoxia/reoxygenation group was 70.55±7.22, higher than those of the control group (14.10±5.93), high glucose group (36.59±2.56) and hypoxia/reoxygenation group (39.04±6.016) (all P<0.05). The relative expression levels of p53 protein in the sip53 transfection group and sip53+siGLUT4 transfection group were 0.322±0.147 and 0.391±0.149, respectively, lower than that in the high glucose combined with negative control group (1.002±0.035) (both P<0.05). The relative expression of GLUT4 protein in the sip53 transfection group was 1.871±0.123, higher than that in the negative control group (1.281±0.232) ( P<0.05). The relative expression of GLUT4 protein in the sip53+siGLUT4 transfection group (0.951±0.193) was lower than that in the sip53 transfection group ( P<0.05). The fluorescence intensity of reactive oxygen species in the sip53 transfection group (27.73±0.74) was lower than that in the negative control group (38.83±0.83) ( P<0.05). The fluorescence intensity of reactive oxygen species in the sip53+siGLUT4 transfection group (43.12±5.08) was higher than that in the sip53 transfection group ( P<0.05). The number of mitochondria, the average area of mitochondria, the average perimeter of mitochondria, the number of mitochondrial branches and the average branch length of mitochondria in the sip53 transfection group were (92.27±10.10), (9.25±0.42) μm 2, (10.86±0.58) μm, (83.27±13.57), and (1.81±0.21) μm, respectively. They were higher than (52.36±16.87), (7.44±1.49) μm 2, (9.22±1.11) μm, (52.36±16.87), and (1.22±0.26) μm in the negative control group (all P<0.05). The number of mitochondria, the average area of mitochondria, the average perimeter of mitochondria, the number of mitochondrial branches and the average branch length of mitochondria in the sip53+siGLUT4 transfection group were (53.73±9.49), (6.89±0.61) μm 2, (8.88±0.47) μm, (53.73±9.49), and (1.22±0.17) μm, respectively, lower than those in the sip53 transfection group (all P<0.05). The red-green fluorescence intensity ratio, ATP content, 2-NBDG fluorescence intensity and relative expression of Mfn2 protein in the sip53 transfection group were 1.27±0.23, (0.048±0.021) μmol/10 5 cells, 275 923±10 447 and 2.608±0.581, respectively, higher than those in the negative control group [0.53±0.21, (0.020±0.007) μmol/10 5 cells, 254 875±8 078, and 0.687±0.146, respectively] (all P<0.05). The red-green fluorescence intensity ratio, ATP content, 2-NBDG fluorescence intensity and relative expression of Mfn2 protein in the sip53+siGLUT4 transfection group were 0.40±0.08, (0.011±0.012) μmol/10 5 cells, 199 511±6 855, and 0.649±0.070, respectively, lower than those in the sip53 transfection group (all P<0.05). The relative expression levels of Drp1, Bax/Bcl-2, Caspase-3 proteins and TUNEL fluorescence intensity in the sip53 transfection group were 0.759±0.063, 0.446±0.161, 1.048±0.300, and 48.93±1.48 respectively, lower than those (1.065±0.149, 1.197±0.133, 1.847±0.201, and 67.61±9.99) in the negative control group (all P<0.05). The relative expression levels of Drp1, Bax/Bcl-2, Caspase-3 proteins and TUNEL fluorescence intensity in the sip53+siGLUT4 transfection group were 0.958±0.166, 2.660±0.135, 1.587±0.220, and 63.39±12.84, respectively, higher than those in the sip53 transfection group (all P<0.05). Conclusions:Under the condition of high glucose combined with hypoxia/reoxygenation, p53 induces cardiomyocyte injury by down-regulating GLUT4. Inhibition of p53 can increase the expression of GLUT4, thereby reducing cardiomyocyte injury induced by high glucose combined with hypoxia/reoxygenation.

Neutralizing antibodies have been designed to specifically target and bind to the receptor binding domain (RBD) of spike (S) protein to block severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus from attaching to angiotensin converting enzyme 2 (ACE2). This study reports a distinctive nanobody, designated as VHH21, that directly catalyzes the S-trimer into an irreversible transition state through postfusion conformational changes. Derived from camels immunized with multiple antigens, a set of nanobodies with high affinity for the S1 protein displays abilities to neutralize pseudovirion infections with a broad resistance to variants of concern of SARS-CoV-2, including SARS-CoV and BatRaTG13. Importantly, a super-resolution screening and analysis platform based on visual fluorescence probes was designed and applied to monitor single proteins and protein subunits. A spontaneously occurring dimeric form of VHH21 was obtained to rapidly destroy the S-trimer. Structural analysis via cryogenic electron microscopy revealed that VHH21 targets specific conserved epitopes on the S protein, distinct from the ACE2 binding site on the RBD, which destabilizes the fusion process. This research highlights the potential of VHH21 as an abzyme-like nanobody (nanoabzyme) possessing broad-spectrum binding capabilities and highly effective anti-viral properties and offers a promising strategy for combating coronavirus outbreaks.
Single-Domain Antibodies/immunology* ; Spike Glycoprotein, Coronavirus/metabolism* ; SARS-CoV-2/immunology* ; Animals ; Humans ; Antibodies, Neutralizing/immunology* ; Camelus ; COVID-19/immunology* ; Antibodies, Viral/immunology* ; Angiotensin-Converting Enzyme 2

Purpose To investigate the value of radiomics nomogram for the prediction of p53 abnormal in patient with endometrial carcinoma based on intratumoral and peritumoral MRI.Materials and Methods A total of 145 female patients were pathologically confirmed endometrial carcinoma who underwent pelvic MRI before treatment in Baoding First Central Hospital from January 2020 to April 2024,including 96 patients with p53 wild type and 49 with p53 abnormal.Radiomics features were extracted from both intratumoral and peritumoral regions(2 mm)in diffusion weighted imaging and equilibrium phase of dynamic contrast enhanced MRI,which were selected using least absolute shrinkage and selection operator.Three machine learning algorithms of random forest,K-nearest neighbors and extra trees were conducted to develop the intratumoral,peritumoral and intratumoral combined peritumoral radiomics models.Multivariate Logistic regression was used to constitute the clinical model and nomogram.The performance of these models was evaluated using receiver operating characteristic curve,decision curve analysis and calibration curve.Results The K-nearest neighbors model of the intratumoral combined peritumoral regions performed the best in all radiomics models,the area under the curve were 0.921 and 0.773 in the training cohorts and test cohorts.The radiomics nomogram,which was composed of age,apparent diffusion coefficient and radiomics signatures,achieved the best performance with area under the curve of 0.970 and 0.777 in the training cohorts and test cohorts,respectively.Calibration curve analysis and decision curve analysis demonstrated favorable calibration and clinical utility of the nomogram model.Conclusion The nomogram based on intratumoral and peritumoral MRI radiomics yields a favorable diagnostic value for predicting p53 abnormal in patient with endometrial carcinoma.

Objective To evaluate the performance of a clinical-radiomics model for differentiating focal organizing pneumonia(FOP)and lung adenocarcinoma(LUAD).Methods We retrospectively analyzed the data of 60 patients with FOP confirmed by postoperative pathology at the First Medical Center of the Chinese PLA General Hospital from January,2019 to December,2022,who were matched with 120 LUAD patients using propensity score matching in a 1∶2 ratio.The independent risk factors for FOP were identified by logistic regression analysis of the patients'clinical data.The cohort was divided into a training set(144 patients)and a test set(36 patients)by random sampling.Python 3.7 was used for extracting 1835 features from CT image data of the patients.The radiographic features and clinical data were used to construct the model,whose performance was validated using ROC curves in both the training and test sets.The diagnostic efficacy of the model for FOP and LUAD was evaluated and a diagnostic nomogram was constructed.Results Statistical analysis revealed that an history of was an independent risk factor for FOP(P=0.016),which was correlated with none of the hematological findings(P>0.05).Feature extraction and dimensionality reduction in radiomics yielded 30 significant labels for distinguishing the two diseases.The top 3 most discriminative radiomics labels were GraylevelNonUniformity,SizeZoneNonUniformity and shape-Sphericity.The clinical-radiomics model achieved an AUC of 0.909(95%CI:0.855-0.963)in the training set and 0.901(95%CI:0.803-0.999)in the test set.The model showed a sensitivity of 85.4%,a specificity of 83.5%,and an accuracy of 84.0%in the training set,as compared with 94.7%,70.6%,and 83.3%in the test set,respectively.Conclusion The clinical-radiomics nomogram model shows a good performance for differential diagnosis of FOP and LUAD and may help to minimize misdiagnosis-related overtreatment and improve the patients'outcomes.

Cryoablation therapy with explicit anti-tumor mechanisms and histopathological manifestations has a long history.A large number of clinical practice has shown that cryoablation therapy is safe and effective,making it an ideal tumor treatment method in theory.Previously,its efficacy and clinical application were constrained by the limitations of refrigerants and refrigeration equipment.With the development of the new generation of cryoablation equipment represented by argon helium knives,significant progress has been made in refrigeration efficien-cy,ablation range,and precise temperature measurement,greatly promoting the progression of tumor cryoablation technology.This consensus systematically summarizes the mechanism of cryoablation technology,indications for oral mucosal melanoma(OMM)cryotherapy,clinical treatment process,adverse reactions and management,cryotherapy combination therapy,etc.,aiming to provide reference for carrying out the standardized cryoablation therapy of OMM.

Objective To evaluate the performance of a clinical-radiomics model for differentiating focal organizing pneumonia(FOP)and lung adenocarcinoma(LUAD).Methods We retrospectively analyzed the data of 60 patients with FOP confirmed by postoperative pathology at the First Medical Center of the Chinese PLA General Hospital from January,2019 to December,2022,who were matched with 120 LUAD patients using propensity score matching in a 1∶2 ratio.The independent risk factors for FOP were identified by logistic regression analysis of the patients'clinical data.The cohort was divided into a training set(144 patients)and a test set(36 patients)by random sampling.Python 3.7 was used for extracting 1835 features from CT image data of the patients.The radiographic features and clinical data were used to construct the model,whose performance was validated using ROC curves in both the training and test sets.The diagnostic efficacy of the model for FOP and LUAD was evaluated and a diagnostic nomogram was constructed.Results Statistical analysis revealed that an history of was an independent risk factor for FOP(P=0.016),which was correlated with none of the hematological findings(P>0.05).Feature extraction and dimensionality reduction in radiomics yielded 30 significant labels for distinguishing the two diseases.The top 3 most discriminative radiomics labels were GraylevelNonUniformity,SizeZoneNonUniformity and shape-Sphericity.The clinical-radiomics model achieved an AUC of 0.909(95%CI:0.855-0.963)in the training set and 0.901(95%CI:0.803-0.999)in the test set.The model showed a sensitivity of 85.4%,a specificity of 83.5%,and an accuracy of 84.0%in the training set,as compared with 94.7%,70.6%,and 83.3%in the test set,respectively.Conclusion The clinical-radiomics nomogram model shows a good performance for differential diagnosis of FOP and LUAD and may help to minimize misdiagnosis-related overtreatment and improve the patients'outcomes.

Objective To develop space vegetable cultivation facility(SVCF)and apply to the cultivation of several fruit and vegetable plants on China Space Station.Methods Through conducting experiments and development of regenerative plant cultivation substrate,active water supply,nutrient supply based on controlled-release fertilizer,LED light environment regulation,modular design and development of various functional units of the SVCF,as well as system integration and testing,plant full cycle cultivation verification,the development of the SVCF(also known as Space Vegetable Garden)has been completed.Through the cultivation on-orbit of three vegetables and one fruit plants,the feasibility of cultivation methods with regenerative substrate was tested and the growth characteristics of several plants were studied.Results The facility worked normally,which can supply water actively,detect substrate parameters including water content and conductivity,and adjust the height according to the plant lighting requirement.All performance indicators of the SVCF meet the cultivation requirements of different fruit and vegetable.Three types of lettuce plants("shengcaiwang","hongye","meiluo")and one tomato plants("honglei")were cultivated from seeding to harvest on-orbit.The lettuce and tomato plants grew well.Astronauts participated in the cultivation and management of lettuce and tomato plants,picking and consuming lettuce leaves(about 900 g,fresh weight)and tomato fruit.The tomato seeds were preserved after harvest and returned to earth.The seeds(second generation)was sowed and cultivated on earth.The growth characteristics will be studied and compared with the tomato plants on-orbit.Conclusion The facility performance meets the requirements of several fruit and vegetable plants cultivation on-orbit.The cultivated lettuce and tomato plants grew well.Astronauts picked and consumed lettuce leaves and tomato fruits.The results laid an important technique foundation for the continuous production of multiple types of fruit and vegetable plants on-orbit.