ObjectiveTo explore the effects of different concentrations of oleic acid on human osteosarcoma cell lines 143B and HOS， as well as the impacts of the optimal concentration of oleic acid on cellular lipid droplet synthesis and cell functions. MethodsThe 143B and HOS cells were treated with varying concentrations of oleic acid （0， 25， 50， 100， and 200 µmol/L） for 48 hours. Following treatment， oil red O staining and BODIPY staining were performed to determine the optimal concentration. Subsequently， CCK-8 assays and colony formation experiments were conducted to assess the effect of this optimal concentration of oleic acid on the cell proliferation of both cell lines. Transwell migration assays were utilized to evaluate the influence of the optimal concentration on migratory capacity and Transwell invasion assays were utilized to evaluate the invasive ability. Additionally， Western blot analysis was employed to examine the expression levels of epithelial-mesenchymal transition （EMT） markers Epithelial cadherin （E-cadherin） and Neural cadherin （N-cadherin） in response to treatment with the optimal concentration of oleic acid. ResultsTreatment with oleic acid did not induce significant cell death in either 143B or HOS cells； however， an increase in intracellular lipid droplets was observed alongside enhanced proliferation， migration， invasion capabilities as well as EMT transformation potential （P<0.05）. ConclusionOleic acid induces lipid droplet synthesis in osteosarcoma cells which subsequently promotes their proliferation， migration and invasion abilities along with EMT transformation.

OBJECTIVE To prepare an intelligent responsive liposome-hydrogel nanopreparation co-loaded with gambogic acid （GA）， and characterize its antitumor activity in vitro . METHODS GA-ICG-Lip-gel was prepared by ethanol injection and cold dissolution， incorporating GA and the photosensitizer indocyanine green （ICG）. The appearance and microscopic morphology of GA-ICG-Lip-gel were observed， its encapsulation efficiency and drug loading capacity were measured， and its photothermal conversion performance， photothermal stability， and infrared imaging properties were investigated， along with the determination of its in vitro release profile. Human breast cancer MCF-7 cells were used as objects to investigate the effects of GA-ICG-Lip-gel （or with near-infrared light irradiation） on cell viability， migration ability， and the cellular uptake capacity of GA-ICG-Lip-gel. RESULTS GA-ICG-Lip-gel existed in a solution state at room temperature and transformed into a gel state at 37 ℃. Its microstructure was dense with small pores， and its encapsulation efficiency and drug loading were （96.07±0.86） % and （6.28±1.16） %， respectively. After exposure to near-infrared light， the temperature of GA-ICG-Lip-gel rose above 42 ℃， with no significant attenuation observed in the heating curve. The heating efficiency was dependent on both the irradiation time and drug concentration. Compared to media without gelatinase， the cumulative release rate of GA-ICG-Lip-gel increased in media containing gelatinase. In vitro studies showed that GA-ICG-Lip-gel could be efficiently taken up by MCF-7 cells； GA-ICG-Lip-gel significantly inhibited the viability and migration ability of MCF-7 cells （ P ＜0.05）， and this inhibitory effect was further enhanced under near-infrared light irradiation. CONCLUSIONS This study successfully prepares GA-ICG-Lip-gel， which exhibits favorable photothermal conversion properties and temperature/enzyme dual-responsive drug release characteristics， and demonstrates significant inhibitory effects on the proliferation and migration of breast cancer cells.

The purpose of this study was to clarify the incidence of ketosis in dairy cows in large-scale dairy farms and the effect of ketosis on the subsequent lactation performance of dairy cows.In this experiment,79 perinatal cows in 8 stages of prenatal 21,14,7 days,delivery day,postpartum 3,7,14,21 days were selected to determine the blood biochemical indexes such as BHB,GLU,NE-FA and ALT in their blood.According to the concentration of BHB in the blood,they were divided into healthy group(CON),subclinical ketosis group(SCK)and clinical ketosis group(CK).There was a disorder of mineral elements in dairy cows in the experimental ranch,and the liver function of dairy cows was abnormal,and there was a disorder of energy metabolism.The incidence of keto-sis in dairy cows in the survey pasture was 26.14％,of which the incidence of SCK was 19.32％,and the incidence of CK was 6.82％,which was comparable to the global average incidence and lower than the average incidence in China.Compared with CON cows,SCK and CK cows had energy metabolism disorders and abnormal liver function,and CK cows were more serious.The milk yield of dairy cows with ketosis decreased significantly.The milk fat rate of dairy cows in SCK group was higher than that in CON group,while the milk fat rate of dairy cows in CK group was significantly lower than that in CON group.The fat-to-egg ratio of dairy cows in CK group was sig-nificantly lower than that in SCK group and CON group.The changes of lactation performance in different types of ketosis at 6 months after ketosis were different,and the lactation performance of dairy cows in CON group was the most stable.In the production of pastures,timely and effective treatment and management are essential for restoring the health and milk production performance of dairy cows.Ketosis can lead to disorder of energy metabolism and abnormal liver function in dairy cows,reduce milk yield,and have a continuous effect on lactation performance of dairy cows.Maintaining the health of dairy cows is the key to improving milk production and maintaining sta-ble lactation performance.For different types of ketosis cows,corresponding management and treatment measures should be taken to reduce economic losses.

Objective:To investigate the clinical efficacy of stereotactic body radiotherapy combined with chemotherapy in the treatment of patients with locally advanced pancreatic cancer.Methods:This study is a prospective randomized controlled study that included 80 patients with locally advanced pancreatic cancer admitted to Weinan Central Hospital from January 2020 to June 2022. Patients were randomly divided into two groups using a drawing lots method: the control group ( n = 40) received chemotherapy and conventional medication, while the observation group ( n = 40) received stereotactic body radiotherapy in addition to the treatments in the control group. Each patient was followed up for 1 year. The local control rate of pancreatic cancer, levels of serum carbohydrate antigen 19-9 and carcinoembryonic antigen, abdominal pain relief rate, jaundice resolution rate, incidence of adverse reactions, and 1-year survival rate were compared between the two groups. Results:The local control rate in the observation group reached 85.00% (34/40), which was significantly higher than that in the control group ( χ2 = 5.23, P < 0.05). After treatment, the levels of serum carbohydrate antigen 19-9 and carcinoembryonic antigen in the observation group were significantly lower than those in the control group ( χ2 = 2.60, 3.97, both P < 0.001). The 1-year survival rate in the observation group was 65.00% (26/40), which was significantly higher than that in the control group [42.50% (17/40), χ2 = 4.07, P < 0.05). However, there was no significant difference in incidence of adverse reactions between the two groups ( χ2 = 2.58, 0.05, 0.39, 2.05, all P > 0.05). Conclusions:Stereotactic body radiotherapy combined with chemotherapy has shown good therapeutic effects in patients with locally advanced pancreatic cancer, without significantly increasing the risk of adverse reactions. Therefore, this combination treatment regimen can be considered an effective option for the treatment of locally advanced pancreatic cancer.

Objective:To observe the effect of combining low-frequency transcranial magnetic stimulation (rTMS) with melodic intonation therapy (MIT) in the treatment of aphasia soon after a stroke.Methods:Sixty survivors of a recent stroke with aphasia were randomly divided into a control group, an MIT group and an observation group, each of 20. All received conventional rehabilitation therapy, but for 4 weeks the MIT group was additionally given MIT while the observation group received MIT plus 1Hz rTMS applied over the right inferior frontal gyrus and the posterior superior temporal gyrus. The intensity was 80% of the resting motor threshold. Before and after the treatment, language function and communication ability were evaluated using the Chinese version of the Western Aphasia Battery (WAB) and Communicative Activities of Daily Living (CADL) scale.Results:Significant improvements in the WAB and CADL scores and aphasia quotient (AQ) were observed in both the MIT group and the observation group after the treatment compared to their pre-treatment levels, but the observation group demonstrated significantly higher scores than the other two groups on all three assessments. The MIT group′s average scores were superior to those of the control group.Conclusions:Applying rTMS and MIT in combination significantly improves the speech and the daily communication abilities of aphasics early post-stroke, reducing the severity of their aphasia. The therapy exhibits considerable promise for clinical implementation.

This study aims to clarify the incidence and prevention status of dairy cow mastitis in 79 large-scale pastures in China in 2022,reduce the incidence of dairy cow mastitis,and provide scien-tific basis for formulating prevention and control strategies for dairy cow mastitis in large-scale pastures suitable for China's national conditions.The research team relied on the large-scale ranch of the Comprehensive Experimental Station of National Dairy Industry Technology System to car-ry out research.A total of 79 questionnaires were received.The information on the stock of large-scale farms,the proportion of cows in the herd,the level of cow yields,the incidence and cure rate of cow mastitis,the prevention and control of cow mastitis,diagnosis and treatment programs of cow mastitis,the cost of treating cow mastitis,and the culling rate of farms was collected.Chi-square test and correlation analysis were carried out on the incidence of mastitis in dairy cows and the prevention and control plan,treatment plan,yield level and elimination rate of mastitis in dairy cows by descriptive statistical cross table.The incidence of cow mastitis in some pastures was con-trolled between 5％and 10％,and the cure rate was mostly as high as 98％.According to the co-hort analysis,the importance of prevention and control measures for dairy cow mastitis was as fol-lows:DMT＞DHI＞milk yield＞milk traits＞breast apparent change＞electrical conductivity＞CMT.The importance of treatment measures for dairy cow mastitis is as follows:DMT＞DHI＞milk yield＞milk traits＞breast apparent changes＞electrical conductivity＞CMT.The importance of treatment measures for mastitis is as follows:traditional Chinese medicine＞pathogen detection and drug sensitivity test＞non-steroidal anti-inflammatory drugs＞regular maintenance of milking equipment＞normal milking without treatment＞broad-spectrum antibiotics.The incidence of mas-titis in dairy cows is moderately positively correlated with the yield per unit area,that is,the in-crease in the incidence of mastitis in dairy cows has a certain impact on the yield per unit area of dairy cows.Economic analysis shows that the treatment cost of mastitis is closely related to the in-cidence rate,and the treatment cost of pasture with higher incidence rate is higher.Pastures with higher yields usually have more efficient means of disease prevention and control,and the corre-sponding treatment costs are lower.In conclusion,China's large-scale pastures should strengthen the scientific feeding management level,improve the prevention and control methods to reduce the incidence of cow mastitis,improve the cure rate,and reduce the cost of pasture treatment.

This study was aimed at establishing a novel visual detection method for SARS-CoV-2 based on magnetic beads(MBs)enrichment and Au@Pt nanoparticles(Au@PtNPs)enzyme catalysis.According to the double-antibody sandwich method,MBs conjugated with SARS-CoV-2 antibody 1 enriched SARS-CoV-2 antigen in samples.After magnetic adsorption,separation,and washing,the enriched antigen reacted with Au@PtNPs labeled with SARS-CoV-2 antibody 2,thereby forming an immune com-plex.After another round of magnetic adsorption and washing,the color change was catalyzed by Au@PtNPs in the chromogenic solu-tion,and the results were determined through comparison of color changes:a blue color change indicated a positive result,whereas no obvious color change indicated a negative result.The sensitivity and specificity of this method were evaluated.This study successfully established a visual detection method based on MBs enrichment and Au@PtNPs enzyme catalysis for targeting SARS-CoV-2 antigens.The method was able to detect SARS-CoV-2 antigens within 30 minutes and SARS-CoV-2nucleocapsid protein samples with antigen concentrations above 3.125 ng/mL.The method showed no cross-reactivity with seasonal influenza(H1N1)or influenza B virus nu-cleoprotein.The visual detection method established in this study,based on Au@PtNPs enzyme catalysis and MBs enrichment,has high specificity and sensitivity,is simple and quick to operate,and provides a new approach for rapid viral detection.

Background and Aims:Poorly differentiated thyroid carcinoma(PDTC)is a relatively rare but highly aggressive type of thyroid malignancy.Its biological behavior lies between differentiated and undifferentiated thyroid carcinoma,and it is often characterized by early metastasis,high recurrence rates,and poor survival outcomes.At present,prognostic assessment for PDTC patients primarily relies on traditional indicators such as TNM staging,and there remains a lack of systematic,multi-factorial,and individualized predictive tools.As a visual and quantitative method,the nomogram model has been widely applied in the prognostic evaluation of various tumors;however,its use in PDTC remains limited.This study aims to identify key risk factors associated with poor prognosis in PDTC patients and to construct a risk prediction nomogram model based on multivariate analysis,in order to provide clinical support for individualized postoperative prognostic assessment.Methods:A total of 55 PDTC patients who underwent surgical treatment at our hospital from January 2015 to December 2020 were retrospectively enrolled and followed up for three years.Based on tumor recurrence,metastasis,and mortality during the follow-up period,patients were divided into a good prognosis group and a poor prognosis group.Univariate analysis was performed to screen for clinical features associated with prognosis,followed by multivariate logistic regression to identify independent risk factors.A nomogram risk prediction model was constructed using R software(version 3.5.3),and its predictive performance and calibration were evaluated by receiver operating characteristic(ROC)curve and Bootstrap validation.Results:During the 3-year follow-up,15 patients experienced tumor progression and 1 patient died,resulting in a poor prognosis rate of 29.1%.Univariate analysis showed statistically significant differences in tumor diameter,TNM stage,local invasion,surgical approach,vascular invasion,and nerve involvement between the two groups(all P<0.05).Multivariate logistic regression identified tumor diameter≥3 cm,advanced TNM stage,local invasion,subtotal thyroidectomy,vascular invasion,and nerve involvement as independent risk factors for poor prognosis(all P<0.05).The nomogram model constructed based on these variables demonstrated a C-index of 0.794(95%CI=0.725-0.846),an AUC of 0.817,sensitivity of 82.26%,and specificity of 86.35%,indicating good discriminatory ability and predictive accuracy.Conclusion:Tumor diameter,TNM stage,local invasion,surgical approach,vascular invasion,and nerve involvement are important factors influencing postoperative prognosis in PDTC patients.The nomogram model based on these variables exhibits strong predictive performance and may serve as a valuable tool for individualized risk assessment and therapeutic decision-making in clinical practice.

Objective:To investigate protective effect and mechanism of Tim-3 on sepsis-induced acute lung injury(ALI)by pro-moting mitophagy of alveolar macrophages and inhibiting activation of NLRP3 inflammasome.Methods:LPS-stimulated mouse alveo-lar macrophage(MH-S)model and sepsis-induced ALI mouse model were constructed.Tim-3 siRNA interference technique was used to knock down Tim-3 expression in MH-S cells,and anti-Tim-3 antibody mice were injected intraperitoneally to block Tim-3 function.Western blot was used to detect protein expressions of NLRP3,ASC,cleaved-caspase-1 and mitophagy-related proteins(LC3B,P62,PINK1 and Parkin)in MH-S cells and lung tissue of mice with sepsis-induced ALI.Laser confocal fluorescence staining was used to measure ROS level and mitochondrial membrane potential of MH-S cells.Pathological examination of lung tissue was performed in mice with sepsis-induced ALI in each group,and degree of lung tissue injury was evaluated by Smith scoring system.Bronchoalveolar lavage fluid(BALF)and lung tissue were collected from mice with ALI induced by sepsis in each group.BCA protein quantification method was used to determine protein concentration in BALF.MPO activity in lung tissue was detected by colorimetry.MDA content in lung tissue was detected by TBA method.LC3B protein expression in lung tissue was detected by immunohistochemistry.Results:In mouse alveolar macrophages,Tim-3 knockdown could promote expressions of NLRP3,ASC,cleaved-caspase-1 and P62 proteins,increase ROS release,inhibit PINK1/Parkin pathway activation and LC3B protein expression,and reduce mitochondrial membrane potential.In mice with sepsis-induced ALI,Tim-3 functional blockade could promote expressions of NLRP3,ASC,cleaved-caspase-1 and P62 proteins in lung tissue,aggravate lung pathological injury and pulmonary edema,increase MPO activity and MDA content in lung tissue,and reduce positive rate of LC3B protein.Conclusion:Tim-3 plays a protective role in sepsis-induced ALI by promoting mitophagy in alveolar macrophages and inhibiting NLRP3 inflammasome activation via PINK1/Parkin.

Objective:To explore the spatial correlation between gray matter volume (GMV) changes and neurotransmitter receptors/transporters in patients with first-episode schizophrenia (FES) .Methods:Fifty-four FES patients(FES group) and fifty-nine healthy controls (HC group) were selected from June 2014 to May 2020 in the Psychiatry Department of Renmin Hospital of Wuhan University. Structural magnetic resonance imaging (sMRI) was conducted on all subjects. Differences of GMV were compared across 400 cortical regions and 32 subcortical regions. Based on the positron emission tomography(PET) data from Neuromaps, which provides the density of 19 different neurotransmitter receptors and transporters, Spearman correlation analysis was performed to evaluate the spatial correlation between GMV changes and neurotransmitter systems.Results:Compared to the HC group, FES group exhibited significant GMV reductions in widespread cortical (90/400) and subcortical (6/32) regions (all FDR-corrected P<0.05). The effect size of GMV reduction (Cohen’s d) showed significant positive correlations with the density of 5-hydroxytryptamine 1a(5HT1a) ( r=0.400, Pspin=0.002), γ-aminobutyric acid type A receptor(GABA A)( r=0.307, Pspin=0.002), and metabotropic glutamate receptor 5(mGluR5) ( r=0.275, Pspin=0.020) receptors (all FDR-corrected P<0.05). Conclusion:GMV reductions in a wide range of brain regions existed in patients with FES. There are significant correlations between 5HT1a, GABA A and mGluR5 receptors and gray matter reduction in patients with FES. The disorder of these neurotransmitter receptors may be the potential neurobiological mechanism of gray matter structural abnormalities in the early stage of schizophrenia.