OBJECTIVE To investigate the mechanism by which Shenfukang capsule （SFK） ameliorates renal interstitial fibrosis （RIF） by regulating macrophage-fibroblast crosstalk via the signal transducer and activator of transcription 3 （STAT3）/Yes-associated protein （YAP） pathway. METHODS RAW264.7 macrophages were induced to polarize with transforming growth factor-β 1 （TGF-β 1 ）. Subsequently， a non-contact co-culture system of macrophages with renal fibroblasts NRK-49F， as well as a non-contact co-culture system of macrophages with YAP-knockdown renal fibroblasts， were established. Cells were treated with low， medium， and high concentrations （4， 8， 16 μg/mL） of SFK， as well as a STAT3 inhibitor （STAT3-I，1.64 μg/mL） and losartan potassium tablets （positive control， 0.28 μg/mL）， for 48 h. After intervention， the protein and mRNA expression levels of CD86， CD163， and STAT3 in macrophages， as well as CD86， F4/80， and STAT3 mRNA， were detected. In co-cultured renal fibroblasts， the protein and mRNA expression levels of α -smooth muscle actin （ α -SMA）， Vimentin， collagen type Ⅰ （Col-Ⅰ）， matrix metalloproteinase-1 （MMP-1）， and YAP were detected. In co-cultured YAP-knockdown renal fibroblasts， the protein and mRNA expression levels of α -SMA， Vimentin， and MMP-1 were also detected. RESULTS Following TGF-β 1 induction， the protein expression levels of CD86， CD163， and STAT3， as well as the mRNA expression levels of CD86， F4/80， and STAT3 in macrophages were significantly increased （ P ＜0.05）. The conditioned medium from polarized macrophages activated renal fibroblasts， as evidenced by significantly increased protein and mRNA expression levels of α -SMA， Vimentin， Col-Ⅰ， and YAP， and significantly decreased protein and mRNA expression levels of MMP-1 in renal fibroblasts （ P ＜0.05）. Treatment with SFK and STAT3-I reversed the changes in the above indicators， with the medium concentration SFK group showing stronge r effects on some indicators than the low and high concentration groups. In the non-contact co-culture experiment of macrophages and YAP-knockdown renal fibroblasts， there were no statistically significant differences in the protein and mRNA expression of α-SMA， Vimentin， and MMP-1 among the groups. CONCLUSIONS SFK can inhibit macrophage-renal fibroblast crosstalk by blocking the STAT3/YAP pathway， thereby delaying the progression of RIF.

AIM: To investigate whether proanthocyanidins(PAC)can ameliorate ferroptosis in retinal pigment epithelium(RPE)cells via the Nrf2/GPX4 pathway, and to evaluate its protective effect in dry age-related macular degeneration(ARMD). METHODS: The protective effects of PAC against ferroptosis were evaluated using an in vitro model of H2O2-stimulated human RPE cells(ARPE-19)and an in vivo dry ARMD animal model induced by sodium iodate(NaIO3). CCK-8 assay was applied to assess cell viability. Calcein-AM/PI staining was applied to determine the level of cell death. Lipid peroxidation(LPO)levels were measured using a lipid peroxidation assay kit. CM-H2DCFDA was used to detect the level of reactive oxygen species(ROS)in cells. MitoSOX probe was employed to measure the mitochondrial ROS level. JC-1 staining was used to evaluate the changes in mitochondrial membrane potential. Western Blot was performed to detect the expression levels of proteins(Nrf2, GPX4, and HO-1)related to the Nrf2 pathway. Retinal flat mounts were used to evaluate the structure and function of RPE. Hematoxylin and eosin(H & E)staining was applied to assess the morphological changes in rat retinas. Rhodopsin and opsin staining was used to evaluate visual function in the retina. TUNEL staining was employed to detect apoptosis in retinal cells. RESULTS: H2O2 exacerbates ferroptosis in ARPE-19 cells, as evidenced by elevated levels of Fe2+, ROS, and LPO. PAC preconditioning ameliorates mitochondrial function, reduces intracellular Fe2+, ROS, and LPO levels, and suppresses ferroptosis in RPE cells via activation of the Nrf2/GPX4 signaling pathway. In a NaIO3-induced dry ARMD model, both PAC and the ferroptosis inhibitor Fer-1 reverse the downregulation of retinal Nrf2/GPX4 expression and attenuate retinal cell death.CONCLUSION:PAC inhibits ferroptosis via the Nrf2/GPX4 pathway, offering a novel potential therapeutic strategy for retinal degeneration.

This report presented the first imported case of alveolar echinococcosis in Tianzhu Tibetan Autonomous County, Gansu Province. The patient was a local resident who had worked in Hotan Prefecture, Xinjiang Uygur Autonomous Region from 2019 to 2023, an area co-endemic for cystic and alveolar echinococcosis. He presented with hepatic discomfort in June 2023. Imaging examinations revealed an 8 cm × 6 cm mass in the left liver lobe, and serological testing showed positive IgG antibodies against Echinococcus. Post-surgical pathology confirmed the diagnosis of alveolar echinococcosis. Given that Tianzhu County has historically been endemic only for cystic echinococcosis and has not reported local cases of alveolar echinococcosis, this case was identified as an imported case based on epidemiological history. The case underwent left lateral hepatic lobectomy and no recurrence was seen at 2-year follow-up post-surgery. It is recommended to enhance the identification of different types of echinococcosis among clinicians, improve health education among migrant laborers, reinforce active screening among returning labors, and advance a One Health concept-based, cross-regional “human-animal-environment” joint prevention and control mechanism to prevent the risk of imported echinococcosis.

Objective To establish and internally validate a predictive model for poorly differentiated adenocarcinoma based on CT imaging and tumor marker results. Methods Patients with solid and partially solid lung nodules who underwent lung nodule surgery at the Department of Thoracic Surgery, the Affiliated Brain Hospital of Nanjing Medical University in 2023 were selected and randomly divided into a training set and a validation set at a ratio of 7:3. Patients' CT features, including average density value, maximum diameter, pleural indentation sign, and bronchial inflation sign, as well as patient tumor marker results, were collected. Based on postoperative pathological results, patients were divided into a poorly differentiated adenocarcinoma group and a non-poorly differentiated adenocarcinoma group. Univariate analysis and logistic regression analysis were performed on the training set to establish the predictive model. The receiver operating characteristic (ROC) curve was used to evaluate the model's discriminability, the calibration curve to assess the model's consistency, and the decision curve to evaluate the clinical value of the model, which was then validated in the validation set. Results A total of 299 patients were included, with 103 males and 196 females, with a median age of 57.00 (51.00, 67.25) years. There were 211 patients in the training set and 88 patients in the validation set. Multivariate analysis showed that carcinoembryonic antigen (CEA) value [OR=1.476, 95%CI (1.184, 1.983), P=0.002], cytokeratin 19 fragment antigen (CYFRA21-1) value [OR=1.388, 95%CI (1.084, 1.993), P=0.035], maximum tumor diameter [OR=6.233, 95%CI (1.069, 15.415), P=0.017], and average density [OR=1.083, 95%CI (1.020, 1.194), P=0.040] were independent risk factors for solid and partially solid lung nodules as poorly differentiated adenocarcinoma. Based on this, a predictive model was constructed with an area under the ROC curve of 0.896 [95%CI (0.810, 0.982)], a maximum Youden index corresponding cut-off value of 0.103, sensitivity of 0.750, and specificity of 0.936. Using the Bootstrap method for 1000 samplings, the calibration curve predicted probability was consistent with actual risk. Decision curve analysis indicated positive benefits across all prediction probabilities, demonstrating good clinical value. Conclusion For patients with solid and partially solid lung nodules, preoperative use of CT to measure tumor average density value and maximum diameter, combined with tumor markers CEA and CYFRA21-1 values, can effectively predict whether it is poorly differentiated adenocarcinoma, allowing for early intervention.

BACKGROUND:At present,osteoarthritis has become a major disease affecting the quality of life of the elderly,and the therapeutic effect is poor,often focusing on preventing the disease process,and the pathogenesis of osteoarthritis is still not fully understood.Bioinformatics analysis was carried out to explore the main pathogenesis of osteoarthritis and related mechanisms of gene coding regulation. OBJECTIVE:To screen core differential genes with a major role in osteoarthritis by gene expression profiling. METHODS:Datasets were downloaded from the Gene Expression Omnibus(GEO):GSE114007,GSE117999,and GSE129147.Differential genes in the GSE114007 and GSE117999 data collections were screened using R software,performing differential genes to weighted gene co-expression network analysis.The module genes most relevant to osteoarthritis were selected to perform protein interaction analysis.Candidate core genes were selected using the cytocape software.The candidate core genes were subsequently subjected to least absolute shrinkage and selection operator regression and COX analysis to identify the core genes with a key role in osteoarthritis.The accuracy of the core genes was validated using an external dataset,GSE129147. RESULTS AND CONCLUSION:(1)A total of 477 differential genes were identified,265 differential genes associated with osteoarthritis were obtained by weighted gene co-expression network analysis,and 8 candidate core genes were identified.The least absolute shrinkage and selection operator regression analysis finally yielded a differential gene ASPM with core value that was externally validated.(2)It is concluded that abnormal gene ASPM expression screened by bioinformatics plays a key central role in osteoarthritis.

BACKGROUND:Macrophage migration inhibitory factor(MIF)is a pleiotropic cytokine,which is secreted in different types of stem cells and can regulate the proliferation,differentiation and migration of various types of stem cells.Our previous research has confirmed that human embryonic stem cells secrete MIF and that its concentration in the culture medium is relatively stable.However,whether MIF is involved in the survival,proliferation and differentiation of human embryonic stem cells remains unclear. OBJECTIVE:To investigate the effects of MIF on survival,proliferation,and differentiation of human embryonic stem cells. METHODS:(1)Human embryonic stem cells H9 were cultured.The growth curve of cells was detected and plotted by CCK-8 assay.Enzyme-linked immunosorbent assay was used to determine the level of MIF in the medium.(2)To determine the effects of exogenous MIF on the survival and proliferation of human embryonic stem cells,different groups were established:the control group,which was cultured in stem cell medium without any modifications;the exogenous MIF group,which was treated with different concentrations(30,100,300 ng/mL)of MIF in the stem cell medium;the MIF inhibitor ISO-1 group,which was treated with different concentrations(2,7,21 μmol/L)of ISO-1 in the stem cell medium;and the MIF+ISO-1 group,which was treated with different concentrations of ISO-1 along with 100 ng/mL of MIF.Cell viability was assessed using the CCK-8 assay.(3)To further elucidate the effect of MIF gene on survival and proliferation of human embryonic stem cell,the MIF knockout H9 cell line was constructed by CRISPR-Cas 9 technology to observe the lineage establishment.(4)To determine the effect of high concentrations of MIF on human embryonic stem cell differentiation,100 ng/mL MIF and 100 ng/mL of CXCR4 neutralizing antibody were separately added to the normal stem cell culture medium.The expression levels of self-renewal factors(KLF4,c-MYC,NANOG,OCT4,and SOX2)and differentiation transcription factors(FOXA2,OTX2)were measured using real-time quantitative polymerase chain reaction,immunofluorescence staining,and western blot analysis. RESULTS AND CONCLUSION:(1)The logarithmic growth phase of H9 cells was between 3-6 days.Under normal growth conditions,human embryonic stem cells secreted MIF at a concentration of approximately 20 ng/mL,independent of cell quantity.(2)Compared to the control group,the addition of different concentrations of MIF had no effect on the proliferation of human embryonic stem cells(P>0.05).ISO-1 significantly inhibited the proliferation of human embryonic stem cells,with a stronger inhibition observed at higher concentrations of ISO-1(P<0.05).The addition of MIF in the presence of ISO-1 reduced the inhibitory effect of ISO-1(P<0.05).(3)Real-time quantitative polymerase chain reaction showed that knocking out 50%of the MIF gene resulted in a significant decrease in the growth vitality of human embryonic stem cells and failure to establish cell lines.(4)Adding 100 ng/mL exogenous MIF to the culture medium resulted in a decrease in the mRNA,protein,and fluorescence expression levels of the self-renewal transcription factor KLF4,while the mRNA,protein,and fluorescence expression levels of the differentiation factor FOXA2 increased.(5)When 100 ng/mL CXCR4 neutralizing antibody was added to the culture medium,the mRNA and protein expression levels of KLF4 increased,while the mRNA and protein expression levels of FOXA2 decreased,contrary to the expression trend observed in the MIF group.In conclusion,the endogenous secretion of MIF by human embryonic stem cells is essential for their survival.The addition of MIF to the culture medium does not promote the proliferation of human embryonic stem cells.However,it can lead to a decrease in the expression of the self-renewal factor KLF4 and an increase in the expression of the transcription factor FOXA2.This provides a clue for further investigation into the effects and mechanisms of MIF on the differentiation of human embryonic stem cells.The MIF-CXCR4 axis plays a regulatory role in this process.

Objective To observe the effects of Hedysarum Polybotrys polysaccharide(HPS)on the glucose metabolism of small intestinal smooth muscle in rats with diabetic gastroparesis(DGP)of spleen qi deficiency syndrome;To explore its mechanism based on AMPK/GLUT4 signaling pathway.Methods Totally 72 male Wistar rats were randomly divided into 12 in the blank group and the remaining 60 rats were assigned to the model group.The DGP spleen qi deficiency syndrome model was replicated by intraperitoneal injection of streptozotocin+irregular feeding with high-fat and high sugar feed combined with swimming exhaustion method.The model rats were randomly divided into model group,metformin group and HPS high-,medium-and low-dosage groups.The metformin group was given 100 mg/kg metformin hydrochloride by gavage,while the HPS high-,medium-and low-dosage groups were given 200,100 and 50 mg/kg HPS by gavage,respectively.The blank group and model group were given purified water by gavage once a day for 8 consecutive weeks.The gastric emptying rate and small intestine propulsion rate in rats were detected,HE staining was used to observe the smooth muscle morphology of gastric antrum and ileal tissue,immunofluorescence staining was used to detect the expression of glucagon like peptide-1(GLP-1)in ileal tissue,ELISA was used to detect the contents of adiponectin(APN),glucagon(Glu)and insulin(INS)in serum,Western blot was used to detect the protein expressions of glucose transporter(GLUT)4,GLUT1,AMP-activated protein kinase(AMPK)and p-AMPK in ileal tissue.Results Compared with the blank group,the model group rats showed significantly increased random blood glucose,significantly decreased body mass,gastric emptying rate and small intestinal propulsion rate(P<0.01);the edema in the submucosal layer,loose arrangement of connective tissue,infiltration of a small number of lymphocytes,granulocytes and macrophages,reduced number of goblet cells in the ileal tissue,shedding of intestinal villous epithelial cells and widening of the gap between the submucosal layer and the lamina propria;the expression of GLP-1 in ileal tissue was significantly decreased(P<0.05),the contents of serum APN and INS were significantly decreased,and the content of Glu significantly increased(P<0.01),the expressions of GLUT4 and p-AMPK/AMPK proteins in ileal tissue were significantly decreased,while the expression of GLUT1 protein significantly increased(P<0.01).Compared with the model group,rats in metformin group and HPS high-dosage group showed a random decrease in blood glucose,an increase in body mass,gastric emptying rate,and small intestine propulsion rate(P<0.05,P<0.01);a more regular arrangement of gastric antral tissue cells,a reduction of shedding cells and edema,the smooth muscle structure of the ileal tissue was relatively intact,with evenly distributed cells and reduced infiltration of inflammatory cells;the expression of GLP-1 in ileal tissue increased,the contents of serum APN and INS increased,and the content of Glu decreased,the expressions of GLUT4 and p-AMPK/AMPK proteins in ileal tissue increased,while the expression of GLUT1 protein decreased(P<0.05,P<0.01).Conclusion HPS may up-regulate GLUT4 and down-regulate GLUT1 expression through activates AMPK,promotes glucose uptake and utilization by small intestinal smooth muscle,and improves glucose metabolism of DGP rats with spleen qi deficiency syndrome.

AIM To establish a UPLC-MS/MS method for the simultaneous content determination of liquiritin,liquiritin apioside,verbascoside,narirutin,isoacteoside,apigetrin,hesperidin,isoliquiritin,ononin,liquiritigenin,glycyrrhizic acid,isoliquiritigenin,honokiol,obovatol,pogostone and magnolol in Jiawei Huoxiang Zhengqi Soft Capsules.METHODS The analysis was performed on a 40 ℃ thermostatic ZORBAX Eclipse Plus C18 column(2.1 mm×100 mm,1.8 μm),with the mobile phase comprising of 0.1％formic acid-acetonitrile flowing at 0.4 mL/min in a gradient elution manner,and electron spray ionization source was adopted in positive and negative ion scanning with multiple reaction monitoring mode.RESULTS Sixteen constituents showed good linear relationships within their own ranges(r＞0.990 0),whose average recoveries were 83.74％-105.12％with the RSDs of 1.10％-4.8％.CONCLUSION This accurate,sensitive,stable and reproducible method can provide a reference for the overall quality control of Jiawei Huoxiang Zhengqi Soft Capsules.

Objective To observe the effects of Angelica and Astragalus ultrafiltration membrane extracts on Keap1/Nrf2 signaling pathway-related factors in diabetes kidney disease(DKD)model rats;To explore its mechanism in treating oxidative stress injury of DKD.Methods Totally 70 SPF-grade male SD rats were randomly divided into 10 rats for the blank group and the remaining 60 rats were fed with high-sugar and high-fat diet,streptozotocin was intraperitoneally injected after 4 weeks to establish the DKD model.The established model rats were randomly divided into the model group,positive control group(GKT137831 30 mg/kg)and TCM high-,medium-and low-dosage groups(Angelica and Astragalus ultrafiltration membrane extracts 6,3,1.5 g/kg),which were orally administered with corresponding drugs.The blank group and model group were given equivalent volume of distilled water orally once daily for 12 consecutive weeks.Random blood glucose,body mass and 24 h urinary total protein content(24 h-UTP)were detected,biochemical assays were conducted to detect serum creatinine(SCr),blood urea nitrogen(BUN),triglycerides(TG),total cholesterol(TC)and albumin(ALB)contents,HE and Masson staining were used to observe the morphology in renal tissue,the contents of reactive oxygen species(ROS),glutathione(GSH)and malondialdehyde(MDA)were measured by ELISA,the protein expressions of nicotinamide adenine dinucleotide phosphate oxidase 4(Nox4),Kelch-like ECH-associated protein 1(Keap1),nuclear factor erythroid 2-related factor 2(Nrf2),glutamate-cysteine ligase modifier subunit(GCLM)and glutathione peroxidase 4(GPX4)in renal tissue were detected by Western blot.Results Compared with the blank group,the random blood glucose at each time point of model group rats significantly increased(P<0.01),the 24 h-UTP significantly increased(P<0.01),the contents of SCr,BUN and serum TG,TC increased(P<0.01),and ALB content decreased(P<0.01);with loose arrangement and disordered structure of renal tissue,consolidation of glomeruli,shedding of renal tubular epithelial cells,and increased deposition of collagen fibers;the contents of serum ROS and MDA were significantly elevated(P<0.01),while GSH contents decreased(P<0.01);the protein expression of Nox4 and Keap1 in renal tissue significantly increased(P<0.01),the expression of Nrf2,GCLM and GPX4 protein significantly decreased(P<0.01).Compared with the model group,the random blood glucose were significantly reduced at 6,8,10,12 weeks in the treatment groups(P<0.01),24 h-UTP decreased(P<0.01),the contents of SCr,BUN,TG,TC in positive control group and TCM high-,medium-dosage group decreased,with increased ALB content(P<0.01,P<0.05);renal tubular and glomerular lesions were improved,and collagen fiber proliferation decreased;serum ROS and MDA contents were reduced,while GSH content increased(P<0.01,P<0.05);the protein expression of Nox4 and Keap1 in renal tissue decreased(P<0.01),and Nrf2,GCLM and GPX4 protein expression increased(P<0.01).Conclusion Angelica and Astragalus ultrafiltration membrane extracts can effectively alleviate injury of renal tissue in DKD rats,and its mechanism may be related to the regulation of the Keap1/Nrf2 signaling pathway and inhibition of oxidative stress response.