Objective To investigate the effects of a 100 mT static magnetic field(SMF)on emotional behavior and brain damage-related molecules in mice.Methods Fifty-eight C57BL/6N mice were randomly divided into control group(n=25)and observation group(n=33).Mice in observation group were exposed to a 100 mT SMF for 0.5 h/d over 14 consecutive days,while mice in control group underwent pseudo-exposure.On the 7 and 14 days of exposure,anxiety-like behavior was assessed using open field and elevated plus maze tests.Cerebral blood flow was monitored using laser speckle imaging,and the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,IL-4,central nervous system specific protein β(S100β),neuron-specific enolase(NSE),and brain-derived neurotrophic factor(BDNF)were measured by radioimmunoassay.BDNF expression in the brain was detected by immunofluorescence.Results On the 7 and 14 days of SMF exposure,the open field and elevated plus maze tests showed no statistically significant differences between observation and control groups in the frequencies,durations,and distance entering the central area of the open field and the open arm of the elevated plus maze(P>0.05).Laser speckle imaging revealed no significant difference in cerebral cortical perfusion compared with pre-exposure period(P>0.05).The results of radioimmunoassay showed that compared with control group,on the 7 d of SMF exposure,the serum IL-1β,NSE and S100β levels were significantly increased(P<0.05),the serum BDNF level was significantly decreased(P<0.05),and the IL-1β and TNF-α contents in brain tissues were significantly increased in observation group(P<0.01).On the 14 d of SMF exposure,serum IL-1β,TNF-α,NSE,and S100β levels were significantly increased(P<0.05,P<0.0001),and the brain IL-1β and TNF-α levels were significantly increased(P<0.01)in observation group.No statistically significant differences were found in anti-inflammatory cytokine IL-4 level of serum and brain tissue or BDNF content of brain tissue between the two groups(P>0.05).Conclusion Continuous exposure to a 100 mT SMF for 14 d at 0.5 h/d induces neuroinflammation and brain damage in mice,without inducing anxiety-like behavior.

Objective To investigate the value of secreted frizzled-related protein 1(SFRP1),GATA binding protein 3(GATA3)combined with human papillomavirus deoxyribonucleic acid(HPV-DNA)in predicting the risk of postoperative recurrence of high-grade cervical intraepithelial neoplasia(CIN)with positive thin-prep cytologic test(TCT).Methods A total of 200 patients with high-grade CIN and positive TCT results from July 2021 to July 2023 were selected as study subjects.All patients underwent cervical conization,and were divided into recurrence group and non-recurrence group according to the recurrence status one year after surgery.The clinical data,HPV-DNA viral load,and the relative expression levels of SFRP1 mRNA and GATA3 mRNA in cer-vical tissues were compared between the two groups.The HPV-DNA viral load,SFRP1 mRNA,and GATA3 mRNA expression levels in patients with different CIN grades were compared,and their cor-relations with CIN grade and the risk of postoperative recurrence were analyzed.The predictive val-ues of SFRP1,GATA3,and HPV-DNA for the risk of postoperative recurrence were analyzed.Results Among 200 patients,2 were lost during the follow-up period.The recurrence group(n=30)had higher proportions of CIN grade Ⅲ,glandular involvement,positive surgical margins,high-risk human papillomavirus(HPV),higher HPV-DNA viral load,and a higher proportion of HPV positivity at 6 months after surgery compared with the non-recurrence group(n=168),and the differences were statistically significant(P＜0.05).The relative expression levels of SFRP1 mRNA and GATA3 mRNA in cervical tissues of the recurrence group were lower than those of the non-recur-rence group,and the differences were statistically significant(P＜0.001).The HPV-DNA level in pa-tients with CIN grade Ⅲ was higher than that in those with CIN grade Ⅱ,and the relative expres-sion levels of SFRP1 mRNA and GATA3 mRNA in cervical tissues were lower than those in patients with CIN grade Ⅱ,and the differences were all statistically significant(P＜0.001).Spearman cor-relation analysis showed that HPV-DNA was positively correlated with CIN grade(P＜0.001),while the relative expression levels of SFRP1 mRNA and GATA3 mRNA in cervical tissues were neg-atively correlated with CIN grade(P＜0.001).Logistic regression analysis showed that HPV-DNA,the relative expression levels of SFRP1 mRNA and GATA3 mRNA in cervical tissues were independ-ent influencing factors for the risk of postoperative recurrence(P＜0.001).The receiver operating characteristic(ROC)curve results showed that the areas under the curve(AUCs)for predicting the risk of postoperative recurrence by HPV-DNA,the relative expression levels of SFRP1 mRNA and GATA3 mRNA in cervical tissues were 0.787,0.781,and 0.764,respectively,with sensitivities of 80.00％,76.67％,and 76.67％,and specificities of 74.40％,73.81％,and 67.26％,respec-tively.The AUC for the combined prediction of the risk of postoperative recurrence by the three indica-tors was 0.924,with a sensitivity of 80.00％and a specificity of 93.45％,which was superior to the pre-dictive values of each indicator alone(Z=3.159,3.306,4.018,P＜0.001).Conclusion HPV-DNA,SFRP1,and GATA3 in cervical tissues have certain predictive values for the risk of postoperative re-currence in patients with high-grade CIN and positive TCT results,and combined detection can im-prove the predictive efficacy.

Poly(lactic-co-glycolide)(PLGA)is a key excipient in long-acting sustained-release preparations,and its degradation properties directly affect the drug release behavior.In this study,PLGA microspheres were prepared by microfluidic techniques,and the morphology changes of the microspheres were observed by scanning electron microscopy(SEM).In alkaline environment,due to the accelerated hydrolysis of ester bonds,the surface of the microspheres was rapidly dissolved and eroded,and the degradation rate was significantly higher than that in acidic environment.High temperature accelerated the degradation of PLGA microspheres.Under neutral and alkaline conditions,the microspheres showed aggregation and adhesion.Under acidic conditions,the microspheres gradually decomposed into irregular fragments.The high ionic strength further promoted the surface corrosion of the microspheres,especially under extreme pH conditions.Simultaneously,PLGA microspheres encapsulating coumarin were prepared to simulate the microsphere formulation.The release rate of coumarin after degradation of the microspheres under different conditions was observed by measuring the absorbance with ultraviolet-visible spectrophotometry.The results were consistent with those of the blank microspheres.This study revealed that the degradation of PLGA microspheres was significantly pH-dependent,temperature sensitive and ion strength responsive.These findings not only helped to understand and optimize the long-term stability and controlled release performance of drug-carrying microspheres,but also provided a theoretical basis for further improvement of PLGA-based drug carrier design.

ObjectiveGliomas in the motor functional area can damage the corticospinal tract (CST), leading to motor dysfunction. Currently, there is a lack of unified methods for evaluating the extent of CST damage, especially in patients with high surgical risk where the minimum distance from the lesion to the CST is less than 10 mm. This study aims to further clarify the classification method and clinical significance of CST morphological changes in these patients. MethodsThis retrospective study analyzed 109 high-risk functional area glioma patients who underwent neurosurgical treatment with preoperative diffusion tensor imaging (DTI) imaging and intraoperative neurostimulation guidance between 2014 and 2024. All patients had a lesion-to-tract distance (LTD) of less than 10 mm between the CST and the lesion. Preoperative DTI evaluation of CST involvement-induced morphological changes were reviewed. Patients were divided into 3 groups: 17 cases (15.6%) with symmetric CST morphology compared to the healthy side (CST symmetry), 48 cases (44.0%) with significant CST morphology changes compared to the healthy side (CST deformation), and 44 cases (40.4%) with CST overlap with the tumor (CST overlap). Then we classified patients according to preoperative assessment of tumor-induced morphological changes, and analyze postoperative motor function for each category. ResultsPostoperative pathology showed a significantly higher proportion of high-grade gliomas (HGG) in the CST overlap group compared to the other two groups (P=0.001). Logistic regression analysis showed that CST overlap was a predictor of HGG (P=0.000). The rate of total tumor resection in the CST deformation group and overlap group was lower than in the CST symmetric group (P=0.008). There was a total of 41 postoperative hemiplegic patients, with 4 cases (23.5%) in the CST symmetric group, 11 cases (22.9%) in the CST deformation group, and 26 cases (59.1%) in the CST overlap group. CST overlap with the tumor predicted postoperative hemiplegia (P=0.016). Two-way ANOVA analysis of the affected/healthy side and CST morphology groups showed significant main effects of CST grouping and healthy-affected side (P=0.017 and P=0.010), with no significant interaction (P=0.31). The fractional anisotropy (FA) value in the CST overlap group and the affected side was lower. A decrease in the FA value on the affected side predicted postoperative hemiplegia (sensitivity 69.2%, specificity 71.9%). ConclusionWe have established a method to predict postoperative hemiplegia in high-risk motor functional area glioma patients based on preoperative CST morphological changes. CST overlap leads to a decrease in CST FA values. This method can be used for precise patient management and aid in accurate preoperative surgical planning.

The small-molecule alkaloid halofuginone(HF)is obtained from febrifugine.Recent studies on HF have aroused widespread attention owing to its universal range of noteworthy biological activities and therapeutic functions,which range from parasite infections and fibrosis to autoimmune diseases.In particular,HF is believed to play an excellent anticancer role by suppressing the proliferation,adhesion,metastasis,and invasion of cancers.This review supports the goal of demonstrating various anticancer effects and molecular mechanisms of HF.In the studies covered in this review,the anticancer molecular mechanisms of HF mainly included transforming growth factor-β(TGF-β)/Smad-3/nuclear factor erythroid 2-related factor 2(Nrf2),serine/threonine kinase proteins(Akt)/mechanistic target of rapa-mycin complex 1(mTORC1)/wingless/integrated(Wnt)/β-catenin,the exosomal microRNA-31(miR-31)/histone deacetylase 2(HDAC2)signaling pathway,and the interaction of the extracellular matrix(ECM)and immune cells.Notably,HF,as a novel type of adenosine triphosphate(ATP)-dependent inhibitor that is often combined with prolyl transfer RNA synthetase(ProRS)and amino acid starvation therapy(AAS)to suppress the formation of ribosome,further exerts a significant effect on the tumor microenvironment(TME).Additionally,the combination of HF with other drugs or therapies obtained universal attention.Our results showed that HF has significant potential for clinical cancer treatment.

Objective To analyze the risk of bias during the experimental process and the shortcomings of the research report by evaluating the methodological and reporting quality of animal experimental studies on the prevention and treatment of precancerous lesions of gastric cancer(PLGC)using TCM compounds.To provide reference for improving the quality of animal experimental research on the prevention and treatment of PLGC with TCM compounds.Methods Experimental literature about the prevention and treatment of PLGC with TCM compounds was retrieved from CNKI,Wanfang Data,VIP,CBM,PubMed,Cochrane Library,Web of science and Embase from January 1,2014 to February 23,2024.SYRCLE assessment tool and ARRIVE 2.0 guideline were used to score the included literature and calculate the"low-risk"compliance rate for each item.Results Totally 213 articles were finally included,including 189 Chinese articles and 24 English articles.The SYRCLE tool score was(12.86±1.29)points,and the"low risk"compliance rate was 32.79％.The score of the necessary items of the ARRIVE 2.0 guideline was(24.15±2.80)points,and the"low risk"compliance rate was 49.08％;the score of the recommended items was(11.28±3.40)points,and the"low risk"compliance rate was 30.27％.In the SYRCLE tool evaluation,144(67.61％)studies did not elaborate on the method of generating the allocation sequence.All studies did not describe the adequacy of allocation concealment and the blinding method in the implementation of bias.Only 51 studies(23.94％)explicitly proposed the success criteria for PLGC modeling,only 66 studies(30.96％)provided detailed information on the statistical methods used,29 studies(13.62％)provided complete ethical statements,and 22 studies(10.33％)reported conflicts of interest.Conclusion There are many problems in the methodological quality and reporting quality of animal experimental literature on the prevention and treatment of PLGC with TCM compounds published from 2014 to 2024,especially the implementation of the random blinding strategy during the experimental process,the calculation details of the sample size,and the reporting of inclusion and exclusion criteria,etc.There are many deficiencies in this aspect.It is recommended to refer to the SYRCLE evaluation tool and the ARRIVE 2.0 guideline list to design and report the research plan,thereby improving the credibility and standardization of the PLGC animal experimental research results.

Objective:To evaluate the role of nemo-like kinase (NLK) in lipopolysaccharide (LPS)-induced inflammatory responses of mouse bone marrow-derived macrophages (BMDMs) and the relationship with glycolysis.Methods:BMDMs were extracted from the wild type (WT) and NLK knockout (NLK -/-) C57BL/6 mice of either sex, aged 6-8 weeks, weighing 20-25 g, were used in this study. After induction, differentiation, maturation and purity identification, BMDMs were divided into phosphate buffer solution (PBS) control group (WT+ PBS group, NLK -/-+ PBS group, n=18) and LPS group (WT+ LPS group, NLK -/-+ LPS group, n=18) by a random number table method. LPS at a final concentration of 1 μg/ml was added to stimulate mature BMDMs for 6 h in LPS group, while the equal volume of PBS was given instead in PBS group. The cell viability was measured by Calcein/PI staining, and the expression of cellular NLK, inflammatory factors (tumor necrosis factor-alpha [TNF-α], interleukin-1 beta [IL-1β] and IL-6) and key glycolytic enzymes (hexokinase 2 [HK2], pyruvate kinase M2 (PKM2) and lactate dehydrogenase A [LDHA])mRNA was detected by real-time fluorescent quantitative polymerase chain reaction. Western blot was used to detect the expression of NLK, HK2, PKM2 and LDHA. The concentrations of TNF-α, IL-1β and IL-6 in the supernatant were determined by enzyme-linked immunosorbent assay. Glucose uptake and lactic acid production were measured by hexokinase method and colorimetric method respectively. Results:Compared with WT-type BMDMs, the expression of NLK protein and mRNA in NLK -/--type BMDMs was significantly down-regulated ( P<0.05). Compared with WT+ PBS group, the BMDM viability was significantly decreased, the expression of TNF-α, IL-1β and IL-6 was up-regulated, the concentrations of TNF-α, IL-1β and IL-6 in the supernatant were increased, the expression of NLK, HK2, PKM2 and LDHA protein and mRNA was up-regulated, and the glucose uptake and lactic acid production were increased in WT+ LPS group ( P<0.05). Compared with WT+ LPS group, the BMDM viability was significantly increased, the expression of TNF-α, IL-1β and IL-6 was down-regulated, the concentrations of TNF-α, IL-1β and IL-6 in the supernatant were decreased, the expression of NLK, HK2, PKM2 and LDHA protein and mRNA was down-regulated, and the glucose uptake and lactic acid production were decreased in NLK -/-+ LPS group ( P<0.05). Conclusions:NLK may enhance LPS-induced inflammatory responses of BMDMs by promoting glycolysis in BMDMs of mice.

To establish a highly specific,sensitive,and efficient method for detecting antibodies a-gainst the Erns protein of classical swine fever virus(CSFV),and to distinguish CSFV vaccine strains from wild strains infections in combination with the E2 subunit vaccine.The purified Erns protein of the CSFV expressed by baculovirus was conjugated to carboxylated magnetic beads as a solid-phase carrier and horseradish peroxidase(HRP),separately.A double-antigen sandwich chemiluminescent enzyme immunoassay(CLEIA)was developed by optimizing various reaction parameters using a fully automated chemiluminescence analyzer.This method was then applied to quantitatively detect Erns protein antibodies in sera from pigs infected with prevalent strains and those immunized with the CSFV E2 subunit vaccine and challenged with field strains.The results showed that the optimal conditions for coupling protein-to-magnetic bead were as follows:coupling buffer pH of 8.0,a protein coupling amount of 2.5 mg/g,blocking solution of 10％BSA,serum sample volume of 20 μL.The optimal dilution of enzyme-labeled antigen was at 1:500 with a one-step reaction time of 15 minutes.The cutoff value of the established CLEIA method for detecting CSFV Erns protein antibodies was 5.83 U/mL and a diagnostic sensitivity of 1:128.No cross-reac-tivity was observed with positive sera against African swine fever virus,pseudorabies virus,porcine circovirus type 2,porcine epidemic diarrhea virus,porcine reproductive and respiratory syndrome virus,or porcine gastroenteritis virus.Additionally,the method yielded negative results with sera from pigs immunized with the E2 subunit vaccine.In repeatability tests,the intra-assay coefficient of variation(CV)ranged from 0.77％to 11.56％,and the inter-assay CV ranged from 10.30％to 14.55％,both below 15％.The positive and negative concordance rates with a commercial CSFV Erns protein antibody detection kit were 95.24％and 92.71％,separately,with an overall concord-ance rate of 93.23％.The double-antigen sandwich chemiluminescence method established in this study exhibits high sensitivity,excellent repeatability,and suitability for automated detection,making it applicable for serological differentiation between CSFV E2 subunit vaccination and infec-tion with prevalent strains.

PURPOSE: To investigate the protective effect of sub-hypothermic mechanical perfusion combined with membrane lung oxygenation on ischemic hypoxic injury of yorkshire brain tissue caused by traumatic blood loss. METHODS: This article performed a random controlled trial. Brain tissue of 7 yorkshire was selected and divided into the sub-low temperature anterograde machine perfusion group (n = 4) and the blank control group (n = 3) using the random number table method. A yorkshire model of brain tissue injury induced by traumatic blood loss was established. Firstly, the perfusion temperature and blood oxygen saturation were monitored in real-time during the perfusion process. The number of red blood cells, hemoglobin content, NA⁺, K⁺, and Ca²⁺ ions concentrations and pH of the perfusate were detected. Following perfusion, we specifically examined the parietal lobe to assess its water content. The prefrontal cortex and hippocampus were then dissected for histological evaluation, allowing us to investigate potential regional differences in tissue injury. The blank control group was sampled directly before perfusion. All statistical analyses and graphs were performed using GraphPad Prism 8.0 Student t-test. All tests were two-sided, and p value of less than 0.05 was considered to indicate statistical significance. RESULTS: The contents of red blood cells and hemoglobin during perfusion were maintained at normal levels but more red blood cells were destroyed 3 h after the perfusion. The blood oxygen saturation of the perfusion group was maintained at 95% - 98%. NA⁺ and K⁺ concentrations were normal most of the time during perfusion but increased significantly at about 4 h. The Ca²⁺ concentration remained within the normal range at each period. Glucose levels were slightly higher than the baseline level. The pH of the perfusion solution was slightly lower at the beginning of perfusion, and then gradually increased to the normal level. The water content of brain tissue in the sub-low and docile perfusion group was 78.95% ± 0.39%, which was significantly higher than that in the control group (75.27% ± 0.55%, t = 10.49, p < 0.001), and the difference was statistically significant. Compared with the blank control group, the structure and morphology of pyramidal neurons in the prefrontal cortex and CA1 region of the hippocampal gyrus were similar, and their integrity was better. The structural integrity of granulosa neurons was destroyed and cell edema increased in the perfusion group compared with the blank control group. Immunofluorescence staining for glail fibrillary acidic protein and Iba1, markers of glial cells, revealed well-preserved cell structures in the perfusion group. While there were indications of abnormal cellular activity, the analysis showed no significant difference in axon thickness or integrity compared to the 1-h blank control group. CONCLUSIONS Mild hypothermic machine perfusion can improve ischemia and hypoxia injury of yorkshire brain tissue caused by traumatic blood loss and delay the necrosis and apoptosis of yorkshire brain tissue by continuous oxygen supply, maintaining ion homeostasis and reducing tissue metabolism level.
Animals ; Perfusion/methods* ; Disease Models, Animal ; Brain Injuries/etiology* ; Swine ; Male ; Hypothermia, Induced/methods*

AIM To investigate the protective effect of Sanfeng Tongqiao Dropping Pills against house dust mite(HDM)-induced allergic asthma in mice.METHODS Compared to the intact BALB/c mice in the blank control group,the BALB/c mice randomly assigned into the model group,the dexamethasone group(0.67 mg/kg),and the low-dose,medium-dose,and high-dose Sanfeng Tongqiao Dropping Pills groups(15,30 and 60 mg/kg),were induced into acute allergic asthma models via weekly intraperitoneal sensitization with 0.1 mL HDM solution(0.5 mg/mL)for three weeks followed by three consecutive daily intranasal challenges with 10 μL HDM solution(2.5 mg/mL)starting in the third week.The drug administered continuously 7 days after the last excitation.The mice had their airway reactive Penh value detected,their pulmonary pathological changes observed by HE staining,their blood eosinophils(EOS)counted,their Th2 cytokines in lung tissue and serum IgE levels detected by ELISA,and their number of peripheral blood mononuclear cells(PBMC)and pulmonary Th2 cells detected by flow cytometry.Chronic allergic asthma was induced in grouped BALB/c mice through repeated intranasal challenges with 10 μL HDM solution(2.5 mg/mL)administered five times weekly for five consecutive weeks.Drug treatment continued for 14 days following the final challenge.After the final treatment,the mice had their pulmonary pathological changes observed by HE staining,and their levels of Th2 cytokines in B ALF and lung tissue and serum IgE detected by ELISA.RESULTS Compared to the blank control group,the acute allergic asthma model group exhibited increases in Penh value,EOS count and IgE level in serum,IL-4 and IL-5 levels in lung tissue(P＜0.01);obvious pulmonary inflammatory cells infiltration,and thickened airway wall;and increase in pulmonary number of Th2 cells(P＜0.01).Compared to the model group,the groups intervened with Sanfeng Tongqiao Dropping Pills demonstrated decreased Penh value,serum EOS count,IgE level and IL-5 level in lung tissue(P＜0.05,P＜0.01);reduced pulmonary inflammatory infiltration and alleviated airway wall thickening;and decreased number of pulmonary Th2 cells.Compared to the blank group,the chronic allergic asthma model group showed obvious pulmonary inflammatory infiltration and airway wall thickening;and increased EOS count and IgE level in serum,IL-4 and IL-13 in lung tissue and IL-14 in BALF(P＜0.05,P＜0.01).Compared to the model group,the groups intervened with either medium-dose or high-dose Sanfeng Tongqiao Dropping Pills demonstrated reduced pulmonary inflammatory infiltration;and decreased serum EOS count,IgE level,IL-13 in lung tissue and IL-14 in BALF(P＜0.05,P＜0.01).CONCLUSION Sanfeng Tongqiao Dropping Pills reduce Th2 cells in peripheral blood and lung tissue,suppress type 2 inflammation,and thereby alleviate allergic asthma.