ObjectiveTo investigate the effect of folic acid-modified crebanine polyethylene glycol-polylactic acid hydroxyacetic acid copolymer（PEG-PLGA） nanoparticles（FA-Cre@PEG-PLGA NPs， hereinafter referred to as NPs） combined with ultrasonic irradiation on subcutaneous tumor of liver cancer in Kunming（KM） mice. MethodsEighty-four healthy male KM mice were utilized to establish a subcutaneous tumor model of mouse hepatocellular carcinoma with H22 cells， then mice were randomly divided into model group， placebo group， hydroxycamptothecin group（8 mg∙kg^-1）， low， medium and high dose crebanine raw material groups（2， 2.5， 3 mg∙kg^-1， hereinafter referred to as the low， medium and high dose crebanine groups， respectively）， low， medium and high dose NPs groups（2， 2.5， 3 mg∙kg^-1）， and low， medium and high dose NPs combined with ultrasonic irradiation groups（2， 2.5， 3 mg∙kg^-1， hereinafter referred to as the low， medium and high dose combination groups， respectively）. The corresponding doses of drugs were administered via tail vein injection， the model group received no treatment， while the placebo group was injected with an equivalent amount of normal saline. Dosing was conducted for a total of 10 times on alternate days. The body mass of the mice was monitored， and parameters such as body mass change rate， thymus index， spleen index， tumor volume， tumor weight， relative tumor growth rate（T/C）， and tumor inhibition rate（TGI） were calculated. Pathological changes in liver and kidney tissues as well as the tumor were observed by hematoxylin-eosin（HE） staining. Additionally， the levels of aspartate aminotransferase（AST）， alanine aminotransferase（ALT）， blood urea nitrogen（BUN） and creatinine（CREA） in serum of mice were detected by biochemical method. Furthermore， the effect of ultrasound on the distribution of NPs in subcutaneous tumors of mouse hepatocellular carcinoma was observed by in vivo imaging technique. ResultsAmong different treatment methods， the combination of NPs and ultrasound irradiation had the best therapeutic effect. Compared with the model group， the body mass growth rates of mice in the medium and high combination groups decreased， while the thymus index and spleen index increased， but there was no statistically significant difference in serum AST， ALT， BUN and CREA levels， indicating that NPs combined with ultrasound irradiation had little effect on the normal physiological state of the body， oth groups had TGI>40% and T/C<60%， indicating a clear anti-tumor effect. Pathological analysis showed that compared with the NPs groups， the combination groups exhibited varying degrees of necrosis in tumor cells， accompanied by less damage to the liver and kidneys. In vivo imaging of small animals showed that compared with the high dose NPs group， the high dose combination group had stronger tumor targeting ability（P<0.01）. ConclusionNPs combined with ultrasonic irradiation can not only effectively targeted the drug to the tumor site， inhibit the subcutaneous tumor growth of mouse liver cancer， but also decrease damage to liver and kidney tissues.

OBJECTIVE To investigate the targeting effect of folic acid-modified crebanine nanoparticles （FA-Cre@PEG- PLGA NPs， hereinafter referred to as “NPs”） combined with ultrasound irradiation on M109 cells in vitro and in vivo after administration， and explore the anti-tumor mechanism. METHODS CCK-8 assay was used to detect the inhibitory effect of NPs combined with ultrasound irradiation on the proliferation of M109 cells， and the best ultrasound time was selected. Using human lung cancer A549 cells as a control， the targeting of NPs combined with ultrasound irradiation to M109 cells was evaluated by free folic acid blocking assay and cell uptake assay. The effects of NPs combined with ultrasound irradiation on the migration， invasion， apoptosis， cell cycle and reactive oxygen species （ROS） levels of M109 cells were detected by cell scratch test， Transwell chamber test and flow cytometry at 1 h after 958401536@qq.com administration； the changes of mitochondrial membrane potential （MMP） were observed by fluorescence inverted microscope. A mouse subcutaneous tumor model of M109 cells was constructed， and the in vivo tumor targeting of NPs combined with ultrasound irradiation was investigated by small animal in vivo imaging technology. RESULTS NPs combined with ultrasound irradiation could significantly inhibit the proliferation of M109 cells， and the optimal ultrasound time was 1 h after administration. The free folic acid could antagonize the inhibitory effect of NPs on the proliferation of M109 cells， and combined with ultrasound irradiation could partially reverse this antagonism. Compared with A549 cells， the uptake rate of NPs in M109 cells was significantly higher （P＜0.01）， and ultrasound irradiation could promote cellular uptake. NPs combined with ultrasound irradiation could inhibit the migration and invasion of M109 cells and block the cell cycle in the G0/G1 and G2/M phases. Compared with control group， the apoptosis rate of M109 cells and ROS level were increased significantly （P＜0.01）， while the MMP decreased significantly （P＜0.01） in the different concentration （100， 200， 300 μg/mL） groups of M109 cells. Compared with the mice in non-ultrasound group， the fluorescence intensity and tumor-targeting index of the tumor site in the 0 h ultrasound group were significantly enhanced （P＜0.05 or P＜0.01）. CONCLUSIONS NPs combined with ultrasound irradiation have a strong targeting effect on M109 cells in vitro and in vivo， the anti-tumor mechanism includes inhibiting cell migration and invasion， blocking cell cycle， and inducing apoptosis.

Objective To investigate the suitable habitats of Phlebotomus chinensis in Gansu Province, so as provide insights into effective management of mountain-type zoonotic visceral leishmaniasis (MT-ZVL). Methods The geographical coordinates of locations where MT-ZVL cases were reported were retrieved in Gansu Province from 2015 to 2023, and data pertaining to 26 environmental variables were captured, including 19 climatic variables (annual mean temperature, mean diurnal range, isothermality, temperature seasonality, maximum temperature of the warmest month, minimum temperature of the coldest month, temperature annual range, mean temperature of the wettest quarter, mean temperature of the driest quarter, mean temperature of the warmest quarter, mean temperature of the coldest quarter, annual precipitation, precipitation of the wettest month, precipitation of the driest month, precipitation seasonality, precipitation of the wettest quarter, precipitation of the driest quarter, precipitation of the warmest quarter, and precipitation of the coldest quarter), five geographical variables (elevation, annual normalized difference vegetation index, vegetation type, landform type and land use type), and two population and economic variables (population distribution and gross domestic product). Twelve species distribution models were built using the biomod2 package in R project, including surface range envelope (SRE) model, generalized linear model (GLM), generalized additive model (GAM), multivariate adaptive regression splines (MARS) model, generalized boosted model (GBM), classification tree analysis (CTA) model, flexible discriminant analysis (FDA) model, maximum entropy (MaxEnt) model, optimized maximum entropy (MAXNET) model, artificial neural network (ANN) model, random forest (RF) model, and extreme gradient boosting (XGBOOST) model. The performance of 12 models was evaluated using the area under the receiver operating characteristic curve (AUC), true skill statistics (TSS), and Kappa coefficient, and single models with high performance was selected to build the optimal ensemble models. Factors affecting the survival of Ph. chinensis were identified based on climatic, geographical, population and economic variables. In addition, the suitable distribution areas of Ph. chinensis were predicted in Gansu Province under shared socioeconomic pathway 126 (SSP126), SSP370 and SSP585 scenarios based on climatic data during the period from 1991 to 2020, from 2041 to 2060 (2050s), and from 2081 to 2100 (2090s) . Results A total of 11 species distribution models were successfully built for prediction of potential distribution areas of Ph. chinensis in Gansu Province, and the RF model had the highest predictive accuracy (AUC = 0.998). The ensemble model built based on the RF model, XGBOOST model, GLM, and MARS model had an increased predictive accuracy (AUC = 0.999) relative to single models. Among the 26 environmental factors, precipitation of the wettest quarter (12.00%), maximum temperature of the warmest month (11.58%), and annual normalized difference vegetation index (11.29%) had the greatest contributions to suitable habitats distribution of Ph. sinensis. Under the climatic conditions from 1991 to 2020, the potential suitable habitat area for Ph. chinensis in Gansu Province was approximately 5.80 × 10⁴ km², of which the highly suitable area was 1.42 × 10⁴ km², and primarily concentrated in the southernmost region of Gansu Province. By the 2050s, the unsuitable and lowly suitable areas for Ph. chinensis in Gansu Province had decreased by varying degrees compared to that of 1991 to 2020 period, while the moderately and highly suitable areas exhibited expansion and migration. By the 2090s, under the SSP126 scenario, the suitable habitat area for Ph. chinensis increased significantly, and under the SSP585 scenario, the highly suitable areas transformed into extremely suitable areas, also showing substantial growth. Future global warming is conducive to the survival and reproduction of Ph. chinensis. From the 2050s to the 2090s, the highly suitable areas for Ph. chinensis in Gansu Province will be projected to expand northward. Under the SSP126 scenario, the suitable habitat area for Ph. chinensis in Gansu Province is expected to increase by 194.75% and 204.79% in the 2050s and 2090s, respectively, compared to that of the 1991 to 2020 period. Under the SSP370 scenario, the moderately and highly suitable areas will be projected to increase by 164.40% and 209.03% in the 2050s and 2090s, respectively, while under the SSP585 scenario, they are expected to increase by 195.98% and 211.66%, respectively. Conclusions The distribution of potential suitable habitats of Ph. sinensis gradually shifts with climatic changes. Intensified surveillance and management of Ph. sinensis is recommended in central and eastern parts of Gansu Province to support early warning of MT-ZVL.

Objective To observe the value of machine learning(ML)models based on CT angiography(CTA)morphology combined with blood inflammation indicators for predicting rupture of intracranial aneurysms(IA).Methods Totally 286 IA patients were enrolled,including 143 cases in ruptured group and 143 case in unruptured group.Among them 200 cases(100 in ruptured subgroup and 100 in unruptured subgroup)were taken as training set,while 86 cases(43 in ruptured subgroup and 43 in unruptured subgroup)were taken as validation set.CTA morphological parameters and blood inflammation indicators were compared between subgroups in training set,and the impact factors of IA ruptured among univariate variables were screened using stepwise logistic regression analysis.Logistic regression(LR),classification and regression tree(CART)and backpropagation neural network(BPNN)models were constructed based on the above impact factors,respectively.Receiver operating characteristic curves were drawn,the area under the curves(AUC)were calculated to evaluate the efficacy of these models for predicting IA rupture.Results Neutrophil,neutrophil-lymphocyte ratio,interleukin-10,tumor necrosis factor-α,transforming growth factor-β,as well as tumor width,height and size ratio(SR)were all impact factors of IA rupture(all P＜0.05),and collinearity diagnosis suggested that no covariate relationship existed among these factors.LR,CART and BPNN models had good efficacy for predicting IA rupture in both training and validation sets with AUC of 0.878-0.993,and BPNN model had the best predictive efficacy(AUC of 0.993,0.976).Conclusion ML models based on CTA morphology combined with blood inflammation indicators could be used to predict IA rupture effectively,among which BPNN model had the best efficacy.

Objective To observe the value of machine learning(ML)models based on CT angiography(CTA)morphology combined with blood inflammation indicators for predicting rupture of intracranial aneurysms(IA).Methods Totally 286 IA patients were enrolled,including 143 cases in ruptured group and 143 case in unruptured group.Among them 200 cases(100 in ruptured subgroup and 100 in unruptured subgroup)were taken as training set,while 86 cases(43 in ruptured subgroup and 43 in unruptured subgroup)were taken as validation set.CTA morphological parameters and blood inflammation indicators were compared between subgroups in training set,and the impact factors of IA ruptured among univariate variables were screened using stepwise logistic regression analysis.Logistic regression(LR),classification and regression tree(CART)and backpropagation neural network(BPNN)models were constructed based on the above impact factors,respectively.Receiver operating characteristic curves were drawn,the area under the curves(AUC)were calculated to evaluate the efficacy of these models for predicting IA rupture.Results Neutrophil,neutrophil-lymphocyte ratio,interleukin-10,tumor necrosis factor-α,transforming growth factor-β,as well as tumor width,height and size ratio(SR)were all impact factors of IA rupture(all P＜0.05),and collinearity diagnosis suggested that no covariate relationship existed among these factors.LR,CART and BPNN models had good efficacy for predicting IA rupture in both training and validation sets with AUC of 0.878-0.993,and BPNN model had the best predictive efficacy(AUC of 0.993,0.976).Conclusion ML models based on CTA morphology combined with blood inflammation indicators could be used to predict IA rupture effectively,among which BPNN model had the best efficacy.

Objective To observe the effects of Xiaomudan Granules on cholesterol synthesis in rats with non-alcoholic fatty liver disease(NAFLD);To explore its mechanism on the treatment of NAFLD based on mTORC1/USP20/HMGCR pathway.Methods Totally 60 SD rats were randomly divided into blank group,model group,Western medicine group(polyene phosphatidylcholine)and TCM low-,medium-and high-dosage groups(Xiaomudan Granules).The blank group was fed with ordinary diet,and the other groups were fed with high-fat diet for 12 weeks to establish NAFLD rat model.After successful modeling,each administration group was given the corresponding drug intragastric administration,and the blank group and model group were given aseptic distilled water intragastric administration for 4 weeks.Body mass and liver mass of rats were recorded,liver index was calculated,and serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C)contents were detected by automatic biochemical analyzer,the morphological changes of liver tissue were observed by HE staining and oil red O staining,real-time fluorescence quantitative PCR and Western blot were used to detect ribosome S6 kinase(S6K),ubiquitin specific protease 20(USP20),3-hydroxy-3-methylglutaryl CoA reduction enzyme(HMGCR)mRNA and p-S6K,S6K,USP20,HMGCR protein expression in liver tissue.Results Compared with the blank group,the body mass,liver mass and liver index of rats in model group significantly increased(P<0.01,P<0.05),the volume of liver lobe increased,the edge was blunted;the contents of serum ALT,AST,TC,TG and LDL-C significantly increased,while HDL-C content significantly decreased(P<0.01);most hepatocytes showed steatosis,significant vacuole and inflammatory infiltration,increased lipid droplets,and significantly increased mRNA expression of USP20 and HMGCR in liver tissue(P<0.01)and protein expressions of p-S6K,USP20 and HMGCR(P<0.01).Compared with the model group,TCM high-dosage group and Western medicine group could significantly decrease body mass,liver mass and liver index of rats(P<0.01,P<0.05),and improve the appearance of liver;decrease the contents of ALT,AST,TC and LDL-C in serum,and increase the content of HDL-C(P<0.01,P<0.05);alleviate hepatocyte steatosis and balloon-like degeneration,reduce lipid droplet deposition,and decrease USP20,HMGCR mRNA and p-S6K,USP20,HMGCR protein expression in liver tissue(P<0.01,P<0.05).Conclusion Xiaomudan Granules may regulate cholesterol synthesis through mTORC1/USP20/HMGCR pathway,and thus play a role in the treatment of NAFLD in rats.

Liver macrophages are important immune cells in the liver,and they express proinflammatory factors and anti-inflammatory factors through polarization into M1 type and M2 type,respectively,thereby playing a role in regulating inflammatory damage response.The malignant transformation of hepatic progenitor cells is the core mechanism of the malignant progression of hepatic precancerous lesions,and its key factor is the continuous stimulation of inflammatory microenvironment,which is closely associated with M1/M2 macrophage polarization.This review mainly focuses on the association between macrophage polarization,chronic inflammation,and malignant transformation of hepatic progenitor cells,so as to provide a theoretical basis for the prevention and treatment of hepatic precancerous lesions.

Objective To explore the otherness of orthotopic injection of cell suspensions and transplantation of tumor tissue blocks to establish orthotopic implantation models of hepatocellular carcinoma in mice,and to provide a technical reference for the establishment of an orthotopic implantation model.Methods Healthy KM mice were divided into four groups:group A,direct injection of H22 cells;group B,direct injection of H22 ascitic cells;group C,transplantation of tissues;and group D,direct injection of saline.Activity and weight changes were observed regularly in each group and survival times were recorded.Liver tumor formation,tumor size,abdominal organ adhesion degree,and metastasis were observed in all groups.B-ultrasound imaging was performed,concentrations of alpha fetoprotein(AFP)and abnormal prothrombin(DCP)were detected,and liver histopathological changes were detected by hematoxylin and eosin staining.Results Mice molding operation time in groups A,B,and C were(3.36±0.44)min,(3.30±0.41)min,and(5.68±0.65)min,respectively.After modeling for 25 days,the rates of model formation in groups A,B,and C were all 100.0%.Severe abdominal adhesions occurred in 40.0%of mice in group A and 60.0%in group B,but in no mice in group C or D.Ascites occurred in 40.0%,100.0%,and 0.0%and abdominal wall tumors in 30.0%,60.0%,and 0.0%of mice in groups A,B,and C,respectively,while 40.0%of mice in group B also had liver metastasis.B-ultrasound imaging,detection of serum AFP and DCP levels,and histopathological result showed smooth liver margins,uneven echo and slightly lower echo mass,maintained high AFP and DCP secretion,and large numbers of inflammatory cells and tumor cells in mice in groups A,B,and C.Conclusions At day 25,all three methods can thus be used to establish orthotopic transplantation models of HCC.Among these,inj ection of cell suspensions demonstrated the advantage of simplicity in operation and the presence of multiple metastatic nodules within the liver,compared to transplantation of tumor tissue.Conversely,transplantation of tumor tissue showed the advantage of causing less impact on the abdomen and other organs when compared to inj ection of cell suspensions.

Objective:To investigate the correlations of urinary Alzheimer-associated neuronal thread protein (AD7c-NTP) with apolipoprotein E (ApoE) allele and cognitive function in patients with subjective cognitive decline (SCD), mild cognitive impairment (MCI) and Alzheimer's disease (AD).Methods:Two hundred and fifty-nine AD patients or patients at prodromal AD stage, including SCD ( n=148), MCI ( n=65), and AD ( n=46), admitted to Department of Neurology, Xuanwu Hospital, Capital Medical University from January 2019 to January 2021, were chosen. Results of neuropsychological assessment, ApoE alleles and urinary AD7c-NTP level were collected in all subjects. Correlations of AD7c-NTP with grading of ApoE alleles and neuropsychological assessment results were analyzed by Spearman's rank correlation. Results:Urinary AD7c-NTP level in AD group was significantly higher than that in MCI group and SCD group ( P<0.05). ApoE genotype test showed that, in these 259 subjects, 24 had ApoE genotype grading 1, 150 had APOE genotype grading 2, 10 had APOE genotype grading 3, and 75 had APOE genotype grading 4; compared with patients with ApoE genotype grading 1 and 2, patients with ApoE genotype grading 4 had significantly increased urinary AD7c-NTP level ( P<0.05). Correlation analysis showed that urine AD7c-NTP level was positively correlated with ApoE genotype grading ( r s=0.264, P<0.001). Urinary AD7c-NTP level was negatively correlated with scores of Montreal cognitive assessment, auditory verbal learning test-N5, animal verbal fluency test and Boston naming test ( P<0.05), and positively correlated with shape trails tests-A and shape trails tests-B scores ( P<0.05). Conclusion:In AD patients or patients at prodromal AD stage, the higher the urinary AD7c-NTP level, the higher the ApoE genotype grading and the greater the decline in cognitive function.

Objective:To investigate the effect of hyperbaric oxygen（HBO）combined with rosuvastatin on the levels of D-dimer（D-D），lipoprotein-associated phospholipase A2（Lp-PLA2），and high-sensitivity C-reactive protein（hs-CRP）in patients with type 2 diabetes mellitus（T2DM）combined with coronary heart disease（CHD）.Method:The clinical data of 167 T2DM patients combined with CHD admitted to Linyi Central Hospital from July 2019 to July 2021 were retrospectively analyzed. They were divided into two groups according to different treatment methods. The control group（ n=78）was given conventional treatment and oral rosuvastatin，and the study group（ n=89）was treated with HBO on the basis of the treatment in the control group. The effective rate，blood viscosity，and the levels of D-D，Lp-PLA2，and hs-CRP were compared between the two groups. Results:After treatment，the effective rate in the study group［93.23%（83/89）］was significantly higher than that of the control group［82.05%（64/78）］，and the difference was statistically significant（ P<0.05 or P<0.01）；the serum levels of D-D，Lp-PLA2，hs-CRP，and blood viscosity in the study group were significantly lower than those of the control group，and the differences were statistically significant（ P<0.05 or P<0.01）. Conclusion:HBO combined with rosuvastatin in the treatment of T2DM patients combined with CHD can be more effective than using rosuvastatin alone in reducing D-D，Lp-PLA2，hs-CRP levels，and blood viscosity，thus achieving better clinical efficacy.