Pulmonary hypertension (PH), marked by sustained elevation of pulmonary artery pressure, imposes a heavy burden on the right ventricle and may culminate in right heart failure. Its pathogenesis is multifaceted, encompassing endothelial dysfunction, vascular smooth muscle proliferation, inflammation, thrombosis, and genetic factors. Animal models serve as core tools for exploring PH mechanisms and therapies, each with unique strengths and limitations. The single-dose monocrotaline (MCT) model is one of the most commonly used experimental animal models of PH and is widely applied in mechanistic studies, drug screening, and efficacy evaluation; it offers simplicity and cost-effectiveness, can induce PH within a short period, yet its pathophysiology differs to some extent from human idiopathic PH. In contrast, the Sugen5416 combined with chronic hypoxia model better mimics PH progression by placing animals under hypoxic conditions to induce pulmonary vasoconstriction and vascular remodeling, but it requires a longer modelling time, and the degree of hypoxia has a substantial impact on experimental outcomes. Beyond these two commonly used modeling approaches, a variety of emerging techniques have been applied in PH research; gene-editing technologies enable precise investigation of specific gene functions in PH. Additionally, induced pluripotent stem cell-based 3D organoid technology allows for individualized modelling while preserving patients' genetic information for precise clinical translation. Each model or technology can simulate different aspects of the pathological processes of human PH, and their findings provide key insights into the nature of the disease and serve as an important platform for the development of novel therapeutic targets. This paper comprehensively describes various animal models and emerging technologies used in PH research, analyzing their characteristics, applications, and limitations, with the aim of providing experimental and technical support for the development of new therapeutic strategies and drugs.

ObjectiveTo investigate the protective effect of α-ketoglutarate （α-KG） on hepatic lipid deposition in offspring caused by arsenic exposure during pregnancy. Methods8-week-old institute of cancer research （ICR） mice were mated in a ratio of 2∶1 between females and males， and the detection of vaginal plugs confirmed pregnant. A total of 32 pregnant mice were randomly divided into four groups： control group， arsenic group， α-KG group， arsenic+α-KG group. On gestational day 0-16 （GD0-GD16）， the arsenic and arsenic+α-KG groups were exposed to sodium arsenite （NaAsO₂ ，15 mg/L） in drinking water everyday， and the α-KG and arsenic+α-KG groups were gavaged with α-KG （2 g/kg） everyday. On GD16， pregnant mice were euthanized to collect fetal liver， and fetal body weight and crown-rump length were measured. Gene expression differences between the control group and the arsenic group were analyzed by transcriptome. The total triglycerides （TGs） and subtypes in fetal liver were detected by liquid chromatography tandem mass spectrometry （LC-MS/MS）. Oil red O staining was used to observe the histopathological changes in the liver. Quantitative polymerase chain reaction （qPCR） was used to detect the expression level of genes related to lipid synthesis， transport， and degradation， and phosphatidylinositol 3' -kinase/ protein kinase B （PI3K/AKT） in the liver of fetus. ResultsTranscriptomics analysis showed that 2 144 genes were downregulated and 1 675 genes were upregulated in the arsenic exposed fetal liver； body weight and crown-rump length were reduced （P_TuKey<0.05）； the level of hepatic TGs was elevated in arsenic group （P_TuKey<0.05）； oil-red O staining showed a significant increase in lipid droplets in arsenic group （P_TuKey<0.01）； the expression of lipid synthesis-related genes were significantly upregulated （P_TuKey<0.05）； the expression of β-oxidation-related genes and lipid degradation-related genes were downregulated （P_TuKey<0.05）； the expression of PI3K， AKT decreased（P_TuKey<0.05）. Compared with the arsenic group， the body weight and crown-rump length of fetus increased in the arsenic+α-KG group （P_TuKey<0.05）； the level of hepatic TGs decreased in the arsenic+α-KG group （P_TuKey<0.05）； oil red O staining showed lipid droplets significantly decreased （P_TuKey<0.01）； the expression of lipid synthesis-related genes were downregulated （P_TuKey<0.05）， the expression of β-oxidation-related genes and lipid degradation-related genes were upregulated （P_TuKey<0.05）； the expression levels of PI3K and AKT increased （P_TuKey<0.05）. Conclusionα-KG alleviated hepatic lipid deposition in offspring exposed to arsenic during pregnancy through activating PI3K/AKT signaling pathway.

ObjectiveHistorically documented Zhejiang Atractylodis Macrocephalae Rhizoma（Baizhu） possesses premium characteristics such as phoenix-like head and crane-like neck， pronounced sweetness， and fragrant aroma. However， its current market circulation is low， and the processed products with Zhejiang-style characteristics are at the risk of being lost. This study aims to preserve the ancient Zhejiang-style processing techniques and evaluate them using modern scientific methods. MethodsMultidimensional intelligent sensory evaluation was used to digitally characterize the "quality-structure" of the external appearance of nine-steamed and nine-processed Baizhu medicinal materials（intermediate processed products） and the "odor-taste" of the internal quality of its decoction pieces（slices）， and the appearance parameters were digitally characterized by colorimeter， texture analyzer， electronic nose and electronic tongue， the chemical composition was analyzed via ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）. Then， cluster analysis on the differences in odor between the medicinal materials（intermediate processed products） and decoction pieces（slices） of nine-steamed and nine-processed Baizhu was conducted， as well as the differences in taste between water-soluble and alcohol-soluble extracts of the decoction pieces（slices）， and the correlation analysis of chroma value-alcohol-soluble extract content-component response value. ResultsThe nine-steamed and nine-processed Baizhu had a dark brown to black epidermis， a brownish-yellow to brownish-gray cross-section， a slightly tough texture， a faint odor， and a slightly sweet， bitter and pungent taste. Texture analyzer measurements revealed minimal adhesion and maximum recovery in the middle section of the characteristic processed Baizhu， consistent with the processing endpoint of thorough steaming and cooking. The head section showed the highest internal hardness， elasticity and chewiness， indicating a denser texture in this area. The electronic nose sensor could clearly distinguish the difference between the medicinal materials and its decoction pieces， with a more significant clustering effect at 60 ℃ for 30 minutes compared to ambient temperature headspace for 2 hours， highlighting the significant impact of the baking degree before slicing on the quality. The electronic tongue taste signal map clearly distinguished the differences between water-soluble and alcohol-soluble extracts of nine-steamed and nine-processed Baizhu decoction pieces， and the addition of auxiliary materials during processing could enhance its alcohol-soluble extract content. A total of 82 chemical components were identified in the characteristic processed Baizhu. After processing， the contents of 58 components increased， while 24 components decreased. Correlation analysis revealed significant negative correlations（P<0.01） between ethanol-soluble extract content and colorimetric values of brightness（L^*）， yellow-bule value（b^*）， and total color difference（E^*_ab）. E^*_ab showed marked negative correlations（P<0.05） with the response values of isochlorogenic acid A and C. ConclusionThis study establishes a modern intelligent sensory evaluation model for multidimensional holographic characterization of nine-steamed and nine-processed Baizhu， clarifying the correlation between increased isochlorogenic acid content and the visual color appearance after different steaming cycles， as well as its intrinsic alcohol-soluble extracts. This provides a reference for quality evaluation and processing standards of the Zhejiang-style characteristic processed products.

ObjectiveTo explore the ameliorative effect of the endoplasmic reticulum stress inhibitor 4-phenylbutyric acid （4-PBA） on bile acid metabolism disorders induced by bacterial lipopolysaccharide （LPS） in late-pregnant mice， and to clarify the regulatory mechanism of endoplasmic reticulum stress （ERS） in intrahepatic cholestasis of pregnancy （ICP）. MethodsTwenty-four pregnant mice were divided into control group， LPS group， 4-PBA group， and LPS+4-PBA group. On the 16th day of pregnancy， the LPS group received intraperitoneal injection of LPS （200 μg/kg） and the 4-PBA group received injection of 4-PBA （150 mg/kg）. In the LPS+4-PBA group， 4-PBA was intraperitoneally injected first， followed by LPS administration 1 h later. Serum total bile acid （TBA）， liver injury markers， bile acid profiles， and the expression of key genes and proteins were detected 6 h after LPS treatment. ResultsLPS exposure increased serum TBA levels in pregnant mice（P<0.05）， activated ERS markers and inflammatory factors， and downregulated bile acid transporters and the expression of the rate limiting enzyme CYP7A1（all P<0.05）. 4-PBA intervention effectively inhibited ERS， reduced TBA levels（P<0.05）， and upregulated Ntcp， Bsep， Mdr3， and Mrp3 gene expression（all P<0.05）. ConclusionERS plays an important role in LPS interference with the homeostasis of bile acid metabolism during pregnancy. Inhibiting ERS can improve bile stasis by regulating bile acid transporters， providing a theoretical basis for targeted therapy of ICP.

Objective To compare the application effects of different puncture techniques in the autonomous arteriovenous fistula puncture of patients undergoing hemodialysis,providing a reference for nurses in selecting puncture methods.Methods A total of 135 patients undergoing hemodialysis at a tertiary hospital in Changzhou from September 2023 to September 2024 were selected and randomly divided into 3 groups,including a multiple puncture group,a rope ladder puncture group,and a buttonhole puncture group.Follow-up was conducted every 3 months,and the first puncture success rate,puncture pain,diameter of blood scab at the puncture site,hemostasis time,and complications of arteriovenous fistula(infection,embolism,aneurysm,stenosis,intimal hyperplasia)were compared among the 3 groups.Results A total of 114 patients completed the study,with 40 in the multiple puncture group,42 in the rope ladder puncture group,and 32 in the buttonhole puncture group.The single-puncture success rate was higher in the multiple single-puncture group(99.4％)and the buttonhole puncture group(99.6％)than it in the rope-ladder puncture group(93.3％),with all P＜0.001.The scab diameter in the buttonhole puncture group(2 mm)was larger than that in the multiple single-puncture group(1 mm)and the rope-ladder puncture group(l mm),with all P＜0.001.The incidence of intimal hyperplasia in the buttonhole puncture group(34.3％)was higher than that in the multiple single-puncture group(10.0％)and the rope-ladder puncture group(11.9％),with all P＜0.05.Conclusion Each of the 3 arteriovenous fistula puncture techniques has its advantages and disadvantages.The multiple puncture group technique can improve the first puncture success rate,and it does not significantly increase the complications of arteriovenous fistula compared to the rope ladder and buttonhole techniques,making it suitable for widespread use.

AIM:To investigate the protective effects of Klotho protein against hypoxia/reoxygenation(H/R)-in-duced damage in rat cardiomyocytes,and to elucidate its underlying mechanisms.METHODS:Rat cardiomyocyte H9c2 cells were divided into 4 groups:control,H/R,low-concentration(1 μmol/L)Klotho+H/R,and high-concentration(10 μmol/L)Klotho+H/R groups.Cells were pretreated with Klotho at specified concentrations before induction of H/R injury.Flow cytometry was used to determine cardiomyocyte apoptosis rates,while reactive oxygen species(ROS)levels were measured using the DCFH-DA probe.Additionally,superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were assessed using biochemical assay kits.Mitochondrial membrane potential was evaluated using the JC-1 as-say,and activity of caspase-3 and caspase-9 was quantified.Western blot analysis was conducted to measure the protein expression of cytochrome C(Cyt-C),B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)in cardio-myocytes from each group.RESULTS:Compared with the control group,the H/R group exhibited significantly increased apoptosis rates(P<0.05),elevated ROS levels and MDA content,decreased SOD activity(P<0.05),reduced mitochon-drial membrane potential(P<0.05),increased caspase-3 and caspase-9 activity(P<0.05),decreased mitochondrial Cyt-C and Bcl-2 protein expression(P<0.05),and increased cytoplasmic Cyt-C and Bax protein expression(P<0.05).In comparison with the H/R group,both low-and high-concentration Klotho treatments significantly reduced cardiomyocyte apoptosis(P<0.05),lowered ROS levels and MDA content(P<0.05),increased SOD activity(P<0.05),restored mito-chondrial membrane potential(P<0.05),decreased caspase-3 and caspase-9 activity(P<0.05),increased mitochondrial Cyt-C and Bcl-2 expression(P<0.05),and decreased cytoplasmic Cyt-C and Bax expression(P<0.05).Notably,the high-concentration Klotho group demonstrated more pronounced protective effects(P<0.05).CONCLUSION:Klotho protein exerts protective effects against H/R-induced cardiomyocyte injury,possibly by inhibiting mitochondria-mediated apoptosis.

Objective:To investigate the role and mechanism of leucine-rich α-2-glycoprotein 1 (LRG1) in the fibrosis of human pterygium fibroblasts (HPFs).Methods:A total of 30 nasal primary pterygium tissues from patients who underwent pterygium excision surgery and 30 nasal normal conjunctival tissues from patients who underwent strabismus correction surgery were collected from the First Affiliated Hospital of Harbin Medical University between January 2022 and March 2023, serving as the pterygium group and normal control group, respectively.LRG1 protein expression in both groups was detected by immunofluorescence staining.The mRNA and protein levels of LRG1 and transforming growth factor-β1 (TGF-β1) were evaluated by quantitative real-time PCR (qRT-PCR) and Western blot.Primary HPFs were cultured from excised pterygium tissues using tissue block adhesion method, and cell morphology was observed.Vmentin and cytokeratin were identified by immunofluorescence staining.HPFs were divided into recombinant human LRG1 (rhLRG1) group and blank control group treated with or without 10 μg/ml rhLRG1 for 24 hours, respectively, and cell migration was evaluated via scratch assay.Additionally, HPFs were divided into blank control group, LRG1 overexpression group and LRG1 knockdown group.HPFs in LRG1 overexpression group and LRG1 knockdown group were transfected with LRG1 overexpression plasmids and small interfering RNA for 24 hours, respectively.TGF-β1 mRNA level was evaluated by qRT-PCR and expression of TGF-β1, fibronectin (FN), type Ⅲ collagen (COL3), and α-smooth muscle actin (α-SMA) proteins were evaluated by Western blot.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of the First Affiliated Hospital of Harbin Medical University (No.2022IIT026).Written informed consent was obtained from each subject.Results:HPFs were successfully isolated, exhibiting spindle-shaped morphology with whorled arrangement, positive identification for vimentin, and negative immunofluorescence staining for cytokeratin.The migration rate of the rhLRG1 group was (83.01±2.56)%, significantly higher than (50.32±4.97)% of the blank control group ( t=9.59, P<0.001).Immunofluorescence staining results showed that compared with normal conjunctival tissue, LRG1 protein was significantly higher expressed in pterygium tissue and was widely distributed in fibrous connective tissue and epithelial layer.Both mRNA and protein levels of LRG1 and TGF-β1 were significantly higher in the pterygium group than in the normal control group (mRNA: t=10.18, 6.15, both P<0.05.protein: t=6.83, 8.79, both P<0.05).In the LRG1 overexpression group, mRNA level of TGF-β1, and protein levels of FN, COL3 and α-SMA were significantly increased compared with the blank control and LRG1 knockdown groups (all P<0.05). Conclusions:LRG1 promotes fibrosis and enhances the migration ability in HPFs, and its mechanism may be associated with the upregulation of the TGF-β1 signaling pathway.

OBJECTIVE To explore the efficacy and safety of sofosbuvir-velpatasvir(SOF/VEL)combined with or without ribavirin(RBV)in treatment of the patients with genotype 3(GT3)chronic hepatitis C(CHC)and liver cirrhosis.METHODS Totally 230 patients with CT3 CHC and liver cirrhosis who were treated in Traditional Chi-nese Hospital of Xinjiang Uygur Autonomous Region,Xinjiang Hetian Specialized Hospital of Infectious Diseases and Xinjiang Manasi County People's Hospital from Jun.2018 to Mar.2023 were recruited as the research sub-jects.The clinical curative effects were observed after the subjects were treated with single SOF-VEL or the com-bination with RBV for 12 to 24 weeks.The indexes including high-sensitivity hepatitis C RNA(HCV RNA),blood routine indexes,liver function indexes and noninvasive diagnosis indexes for liver fibrosis were observed,and the sustained virological response 12 weeks after the treatment(SVR12)was analyzed.RESULTS The mean age of the enrolled patients was(42.31±11.18)years old,the male patients accounted for 66.52％,and there were 137 cases of GT3a and 93 cases of GT3b 93,there were 183 cases of CHC,44 cases of compensated cirrhosis(CC)and 3 cases of decompensated cirrhosis(DCC).There were 189 cases of single HCV infection,33 cases of mixed infections of HCV and HIV,6 cases of mixed infections of HBV/HCV and 2 cases of triple infections of HBV/HCV/HIV.The overall SVR12 of the 230 patients was 99.57％,the SVR12 of the GT3a type patients was 100.00％,the GT3b type patients 98.92％.The SVR12 of the patients with CHC,CC and DCC were 99.45％,100.00％and 100.00％,respectively.The SVR12 of the patients with single HCV infection,HCV/HIV infec-tion,HBV/HCV infection and HBV/HCV/HIV were 99.47％,100.00％,100.00％and 100.00％,respective-ly.No patient quit the direct-acting antivirals(DAAs)treatment due to the drug-induced adverse reactions.1 pa-tient had relapse due to irregular administration of DAAs.CONCLUSION The virological response rate is high a-mong the patients with GT3 CHC and liver cirrhosis who are treated with single SOF/VEL or the combination with RBV,with the safety favorable.

Objective:To establish a method for evaluating the material attributes and skeletal performance of 2208 hydroxypropyl methylcellulose(HPMC),to clarify the influence of different material attributes on the skeletal per-formance of HPMC,to compare the differences between the products of different manufacturers,and to analyze the factors affecting the process.Methods:In this study,the material attributes such as powder chemical properties,viscosity,gelation temperature,thermodynamic properties,weight-average molecular weight,methoxy and hydroxypropoxy contents of different manufacturers were firstly investigated.Then,the water absorption,swelling,and dissolution properties of HPMC blank skeleton tablets were determined using the weighing method.Finally,Principal Component Analysis(PCA)and Orthogonal Partial Least-Squares Discrimination Analysis(OPLS-DA)were used to systematically evaluate the material attributes and skeleton performance of HPMC.The systematic evaluation of each material attribute and skeleton performance of HPMC was carried out to elucidate the intrinsic relationship between each material attribute and skeleton performance of HPMC.Results:The results showed that there were obvious differences in the material attributes of HPMC from different manufacturers,such as the proper-ties of powder,viscosity,and weight-average molecular weight,and that there were differences in the corrosion performance,water absorption,and swelling performance of the skeleton tablets prepared from different manufactur-ers,with the most obvious differences between K4Mand K100M.The results of the PCA and OPLS-DA analyses indicated that these 19 variables showed some correlation with each other.Both mathematical models showed better differentiation and classification effects on HPMC samples,and the OPLS-DA model had better classification effects than the PCA model.Conclusion:Based on the PCA and OPLS-DA models,this study conducted systematical research on HPMC,clarified the degree of influence of different material attributes on the skeletal performance of HPMC,and suggested the addition of HPMC particle size and size distribution,and weight-average molecular weight as the quality standards,which provide a basis for the quality control of the related excipients,the screening of formulation prescriptions,and the improvement of performance.

Objective Exploring alterations in regional homogeneity(ReHo)in the brain of adolescents with depression and suicide attempts.Methods Adolescent patients with depression were included.Those with at least one suicide attempt within one year were classified as the suicide attempt group(32 cases),and those without suicide attempts within one year were classified as the non-suicide attempt group(33 cases).Additionally,a healthy control group(31 individuals)was included.All subjects underwent resting-state functional magnetic resonance imaging(fMRI)scans,and the ReHo values were calculated.The severity of depressive symptoms and the suicide risk were evaluated using the patient health questionnaire-9(PHQ-9)and the mini-international neuropsychiatric interview(MINI)suicide scale,respectively.The ReHo values were compared across groups,and the correlations between ReHo values in distinct brain regions and the PHQ-9 and the MINI suicide scales were analyzed.Results Compared with the healthy control group,the ReHo values in the right and left thalamus,right and left precentral gyrus were decreased in the suicide attempt group(voxel level P<0.001,cluster level P<0.05,GRF corrected),while the ReHo value in the right superior temporal gyrus was increased(voxel level P<0.001,cluster level P<0.05,GRF corrected).Compared with the healthy control group,the ReHo values in the right midbrain and the right thalamus were decreased in the non-suicide attempt group(voxel level P<0.001,cluster level P<0.05,GRF corrected),while the ReHo values in the left middle frontal gyrus,right middle frontal gyrus,and right superior frontal gyrus were increased(voxel level P<0.001,cluster level P<0.05,GRF corrected).Compared with the non-suicide attempt group,the ReHo values in the right medial and paracingulate gyrus were decreased in the suicide attempt group(voxel level P<0.001,cluster level P<0.05,GRF corrected).ReHo values in the right superior temporal gyrus of the suicide attempt group showed a positive correlation with PHQ-9 scores(r=0.407,P=0.026).Conclusion Reduced ReHo values in the right medial and paracingulate gyrus at rest in the group with suicide attempt depression may be a potential neural mechanism for suicide in adolescent depressed patients.