Objective To investigate the effect of berberine and its mechanism in delaying silica-induced lung injury via network pharmacology and molecular docking, followed by verification with animal experiments. Methods i) Targets of berberine and silicosis were obtained from multiple databases, and the common targets were identified. A protein-protein interaction network was constructed using the STRING database, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on the common targets via the DAVID database. Molecular docking was conducted using AutoDock software for validation. ii) Specific pathogen-free Wistar rats were randomly divided into five groups, with six rats in each group. Rats in the model group, pirfenidone group, and low- and high-dose berberine groups received a single non-exposure intratracheal instillation of 1.00 mL silica suspension at a mass concentration of 100 g/L, whereas rats in the control group received an equal volume of 0.9% sodium chloride solution. Rats in the pirfenidone group were administered with pirfenidone solution at a dose of 100 mg/kg body weight, and rats in the low- and high-dose berberine groups were administered with berberine solutions at doses of 100 and 200 mg/kg body weight, respectively. The remaining two groups received equal volumes of 0.9% sodium chloride solution. All treatments were administered by gavage once daily for 28 consecutive days from 24 hours after silica exposure. After intervention, rats in each group were sacrificed. Histopathological changes in lung tissues were observed, and the lung organ coefficient was detected. Serum malondialdehyde (MDA) was detected by colorimetry. Serum inflammatory cytokine were detected by enzyme-linked immunosorbent assay. The expression of fibrosis markers in lung tissues was detected by immunohistochemistry. The protein expression of phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), and phosphorylated AKT (p-AKT) in lung tissues was determined by Western blot. Results i) Network pharmacology analysis identified 46 potential targets. The core targets included AKT1, tumor protein p53, caspase-3, matrix metalloproteinase-9, and albumin. GO enrichment analysis revealed that the targets related to the anti-fibrotic effect of berberine against silicosis were mainly enriched in biological processes including signal transduction, negative regulation of apoptosis, extracellular matrix degradation, and response to reactive oxygen species. KEGG enrichment analysis showed that berberine exerted anti-silicosis effects through multiple signaling pathways, such as the PI3K/AKT pathway, hypoxia-inducible factor-1 pathway, and advanced glycation end product (AGE)-receptor for AGE pathway. ii) Histopathological examination revealed that rats in the model group presented typical pulmonary fibrotic lesions. Pathological lung injury of rats was alleviated in all three intervention groups (pirfenidone group, low- and high-dose berberine group) compared with the model group. The improvement in the high-dose berberine group was superior to that in the low-dose berberine group and was comparable with that in the pirfenidone group. rats in the model group showed increased lung organ coefficient, serum MDA, interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) (all P<0.05), as well as increased expression of typeⅠcollagen (COLⅠ), α-smooth muscle actin (α-SMA) and PI3K, and p-AKT/AKT ratio in lung tissues (all P<0.05) when compared with the control group. The above indicators were lower in rats in both the low- and high-dose berberine groups than those in the model group (all P<0.05). Serum MDA, IL-1β, and TNF-α, as well as relative expression of COLⅠ and α-SMA in lung tissues, were lower in the high-dose berberine group than in the low-dose berberine group (all P<0.05). Conclusion Berberine may alleviate pulmonary inflammation and fibrosis in rats through regulation of the PI3K/AKT signaling pathway, thereby delaying the progression of silicosis.

Aim To observe the effect of lipopolysac-charide(LPS)induced supernatant of BV-2 cells on the inflammatory response and apoptosis of HT22 neu-rons.Methods After the concentration and time of LPS were determined by CCK-8 method,BV-2 cells were cultured with medium without LPS and medium containing LPS,the morphological changes of BV-2 microglia were observed by inverted microscope,and the CD86/CD206 ratio of BV-2 microglia was detected by immunofluorescence.Subsequently,BV-2 cell cul-ture supernatants were isolated and added to HT22 neuronal culture to observe the effect on the inflamma-tory response of HT22 neurons.The proliferation of HT22 neurons was detected by CCK-8 method and EdU method.The structural changes of HT22 neurons were observed under the microscope and examined by urani-um-lead staining.The levels of cytokines interleukin-1β(IL-1β),interleukin-10(IL-10),nuclear factor kappa-B(NF-κB)and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent as-say(Elisa).Neuronal apoptosis was detected by the TUNEL method.The protein expressions of Bax,Bcl-2 and inflammatory factors were detected by Western blot.Results After induction with 1 mg·L-1 LPS,BV-2 cells exhibited increased cell body size,thicker protrusions on both side,and some cells showed de-formed protrusions,the CD86/CD206 ratio in BV-2 cells decreased,promoting the transformation of BV-2 cells from M2 type to M1 type.After treating with the culture supernatant of BV-2 cells,HT22 neuronal cell activity and proliferation were reduced,axons short-ened,and the number of cells decreased.Neuronal cell bodies were enlarged and some cells were de-formed,with damaged cell membranes,round cell nu-clei but displaced nucleoli from the normal position,swollen mitochondria with vacuoles,reduced internal ridge structures,and increased levels of inflammatory factors NF-κB,IL-1 β,and TNF-α(P＜0.05 or P＜0.01),while the anti-inflammatory factor IL-10 de-creased(P＜0.05),protein expression of the pro-apoptotic indicator Bax increased(P＜0.01),and the protein expression of the anti-apoptotic indicator Bcl-2 decreased(P＜0.05).Conclusion After induction of BV-2 cell polarization by LPS,the supernatant could inhibit HT22 neuronal cell viability,upregulate inflam-matory factor expression and promote apoptosis.

Apolipoprotein D(APOD)is a protein that is widely present in animal tissues and is in-volved in the reproductive regulation of the body.In order to investigate the expression regularity of APOD in bactrian camel epididymis and its regulation effect on sperm maturation,this study took the epididymis of bactrian camel during estrus and anestrus as materials,and first cloned the complete sequence of APOD coding sequence(CDS)region.The physicochemical properties of AP-OD were analyzed by ProParam,SOPMA,SWISS-MODEL and MEGA7.0 software.Meanwhile,the expression and distribution of APOD in epididymis were detected by qRT-PCR,Western blot and IHC.The cloning results showed that:the length of the CDS region of APOD gene was 624 bp,encoding 207 amino acids.The APOD sequence of Bactrian camel was highly conserved with the nucleotide and amino acid sequence of alpaca,and the homology of APOD sequence with elk was the lowest.The results of qRT-PCR showed that the mRNA levels of APOD in the head,body and tail of epididymis in estrus were significantly higher than those in estrus(P＜0.01).Western blot results showed that the APOD protein expression and mRNA expression trend was similar in the head and body of the epididymis during anestrus,but the APOD expression level in the tail of the epididymis during anestrus was opposite to the mRNA expression level(P＜0.05).The results of H&E and IHC showed that there were significant differences in epididymal tissue between estrus and anestrus.In addition,APOD showed positive reactions in epididymal epithelial cells,smooth muscle cells,sperm and connective tissue to varying degrees,suggesting that APOD may be in-volved in the maturation of sperm during estrus and anestrus,providing evidence for further explo-ring the regulatory mechanism of APOD's involvement in seasonal estrus.

Objective:To evaluate the impact of Diagnosis-Intervention Packet(DIP)payment reform on medical service costs and efficiency for inpatients in public hospitals,and to compare differences between surgical and medical groups.Methods:A quasi-experimental design was employed,using 605 636 discharged patients from a tertiary hospital in Hebei Province between January 2020 and March 2025 as the sample.The difference-in-differences(DID)model was used to analyze the changes in key indicators between the DIP settlement group(intervention group)and the non-DIP settlement group(control group).Results:Total hospitalization costs,out-of-pocket expenses,and medication costs were significantly reduced in the DIP settlement group(P<0.05),while costs for examinations,nursing,laboratory tests,and treatments increased significantly(P<0.05).Material costs increased by 30.7%in the surgical group(P<0.1)and decreased by 19.8%in the medical group(P<0.01).In terms of efficiency,the average length of stay,time,and cost consumption index all decreased(P<0.01),while the proportion of medical services increased(P<0.01).The case mix index(CMI)showed no significant changes.Conclusion:The DIP reform effectively controlled costs and improved efficiency,but it also resulted in cost shifting and departmental disparities.Therefore,it is necessary to optimize cost control and departmental management policies.

Objective:To investigate the correlation between imaging features of mixed ground-glass nodules(mGGNs)under dual lung enhanced CT and pathological subtypes of lung adenocarcinoma.Methods:Retrospective analysis of clinical data of 102 isolated mGGN lung adenocarcinoma patients admitted to Dalian Central Hospital from October 2016 to October 2018.The patients were divided into adenocarcinoma in situ(AIS)group,minimally invasive adenocarcinoma(MIA)group and invasive adenocarcinoma(IAC)group according to postoperative pathological examination results.Measure the maximum diameter lesions,the maximum diameter of solid components and the proportion of solid components to evaluate imaging features of three groups.The relationship between pathological subtypes of lung adenocarcinoma and baseline features,imaging features,mGGN lesions,maximum diameter of solid components and proportion of solid components were analyzed.The diagnostic value of the maximum diameter of lesions,the maximum diameter of solid components,and the proportion of solid components in IAC were analyzed by receiver operating characteristic(ROC)curves.Results:102 patients were divided into AIS group(n=20),MIA group(n=29)and IAC group(n=53)based on postoperative pathological diagnosis.There was a statistically significant difference in age among the three groups(P＜0.05).Tumor distribution locations:35 cases in the upper lobe of the right lung,10 cases in the middle lobe of the right lung,and 15 cases in the lower lobe of the right lung;23 cases in the upper lobe of the left lung and 19 cases in the lower lobe of the left lung,the tumor distribution locations in the upper lobe of the right lung was relatively high in various pathological subtypes.The lesions in AIS and MIA groups were mostly circular or elliptical in shape,whiile the lesions in the IAC group was mostly irregular in shape.There was a statistically significant difference in morphological comparisons among the three groups(P＜0.05).There was a statistically significant difference in burr sign between MIA group and IAC group(P＜0.05).There was a statistically significant difference in pleural indentation sign and bronchial inflation sign between MIA group and IAC group(P＜0.05).There was a statistically significant difference in the maximum diameter of lesions and the maximum diameter of solid components among the three groups(P＜0.05).The proportion of solid components in IAC group was higher than that in AIS and MIA groups,and the difference was statistically significant(P＜0.05).The ROC curve shows that,the area under curve(AUC)for diagnosing IAC based on the maximum diameter of the lesion,the maximum diameter of the solid component,and the proportion of the solid component were 0.840,0.966 and 0.816,respectively.The AUC of diagnosing IAC with the maximum diameter of the solid component was greater than the AUC of the maximum diameter of the lesion and the proportion of solid components(P＜0.05).Conclusion:Dual lung enhanced CT can evaluate the imaging features of mGGN,and it can distinguish the pathological subtypes of lung adenocarcinoma,when the maximum diameter of the lesion is ≥ 16.5 mm,the maximum diameter of the solid component is ≥5.5 mm,or the proportion of solid component is ≥47.00％,it can effectively diagnose IAC,the maximum diameter of solid components has the best diagnostic efficiency for IAC.

Objective:To compare and observe the visual acuity and ocular anatomical outcome of different subtypes in open-globe injury (OGI) Ⅲ.Methods:A retrospective study. A total of 187 eyes of 187 patients with OGI involving zone Ⅲ who were admitted to the Department of Ophthalmology of The First Affiliated Hospital of Army Medical University from January 2020 to December 2023 were included in the study. According to the 2022 International Globe and Adnexal Trauma Epidemiology Study groups consensus, zone Ⅲ was further divided into Ⅲa zone (5-8 mm posterior to the limbus) and Ⅲb zone (>8 mm posterior to the limbus), with 58 eyes (31%, 58/187) in group Ⅲa and 129 eyes (69%, 129/187) in group Ⅲb. Best corrected visual acuity (BCVA) was examined using the international standard decimal visual acuity chart, converted into the logarithm of the minimum angle of resolution (logMAR) visual acuity when recorded. The injured zone, initial visual acuity, final visual acuity, retinal detachment (RD), uveal prolapse, and proliferative vitreoretinopathy (PVR) were collected. The follow-up time after surgery ≥ 6 months. The final visual acuity and anatomical prognosis of the two groups were observed. Silicone oil dependence, phthisis, and enucleation were defined as poor anatomical outcomes. Multiple linear regression analysis was performed to analyze the impact of zone Ⅲb of OGI on the final visual acuity.Results:At the 6-month follow-up, the logMAR BCVA of group Ⅲa and group Ⅲb was 1.49±1.0 and 2.51±0.85; there was a statistically significant difference in the logMAR BCVA between the two groups ( t=-2.736, P<0.05). Compared with group Ⅲa, the proportion with light perception in group Ⅲb was higher, and the proportions with visual acuity of hand movement, counting fingers, and >0.01 were lower, and the differences were all statistically significant ( P<0.05). Compared with group Ⅲa, RD and PVR were more likely to occur in group Ⅲb, and the differences were all statistically significant ( χ2= 16.696, 8.697; P<0.05). Among the affected eyes in group Ⅲa and group Ⅲb, there were 14 eyes (24.1%, 14/58) and 95 eyes (73.6%, 95/129) with poor final anatomical outcomes respectively; the incidence of poor final anatomical outcomes in group Ⅲb was higher, and the difference was statistically significant ( χ2= 40.332, P<0.01). The results of multiple linear regression analysis showed that initial visual acuity, RD, and uveal prolapse were independent risk factors affecting the final visual acuity (odds ratio=2.407, 4.162, 3.413; P<0.05). Conclusions:Patients with OGI in zone Ⅲb have a worse visual prognosis and a higher incidence of poor anatomical outcomes. The subclassification of zone Ⅲ is helpful for better predicting the prognosis of OGI clinically.

As the world's second largest vector of pathogens,ticks can spread a variety of pathogens by sucking the host's blood.Ticks not only threaten human life and health,but also cause great economic losses in animal husbandry.Artificial breeding of ticks can provide a stable environment for the growth and reproduction of ticks,thereby generating sufficient exper-imental materials for understanding ticks'biological characteristics,studying tick-borne pathogens,and developing anti-tick drugs and vaccines.Current methods of breeding ticks in the laboratory can be roughly divided into two categories:breeding methods using host animals or artificial membranes.The selection of breeding method must be comprehensively considered,ac-cording to tick types,blood-sucking habits,living environments,and other aspects.The development processes of the two methods,and their respective advantages and disadvantages,are described and discussed,to assist laboratories in artificial breeding of ticks.

BACKGROUND:The pathogenesis of inflammatory bowel disease involves inflammation,immune activation,visceral hypersensitivity,and dysbiosis of the gut microbiota.Inflammation promotes the release of inflammatory mediators by immune cells,damaging the enteric nervous system.Enteric glial cells are an important component of the intestinal nervous system and are excellent cells for studying intestinal neuroinflammation.Primary enteric glial cells play a crucial role in exploring cell therapies for intestinal nervous system diseases.Currently,the methods for obtaining these cells are mostly cumbersome.Therefore,finding a convenient and fast method for extracting this cell is crucial.OBJECTIVE:To establish a method for optimizing the isolation,culture,and identification of mouse enteric glial cells.METHODS:0-7-day-old C57BL/6 neonatal mice were euthanized by excessive inhalation of isoflurane.After soaking in 75％alcohol for disinfection,the duodenum(1 cm below the pylorus to 1 cm above the Qu's ligament)was removed by laparotomy at the midline of the abdomen.A 1 mL syringe was filled with DPBS and the intestinal contents were repeatedly rinsed until the intestine became translucent,and the mesentery and blood vessels were peeled off.The duodenum was cut to a size of 1 mm and digested in 0.25％EDTA trypsin for 20 minutes.Then an equal amount of DMEM/F12 complete culture medium was added to terminate digestion.The liquid was filtered through a 100 μm cell filter,centrifuged,and the cells were resuspended in 1 mL of DMEM/F12 complete culture medium.When the cell adhesion growth density reached 80％,cells were digested for subculture.When cells were cultured to the third generation,glial fibrillary acid protein labeled with enteric glial cells was used for identification by immunofluorescence method.RESULTS AND CONCLUSION:The isolated and cultured cells were full of colloids,with protrusions extending outward and passable.Glial fibrillary acid protein staining was positive.This method can successfully isolate and culture enteric glial cells and is easy to operate,providing a stable model for the study of the pathophysiology of the enteric nervous system.

Aim To explore the biological mechanism of ligustilide in the prevention and treatment of osteo-porosis by regulating H-type blood vessels,combined with animal experiments for verification,based on net-work pharmacology and molecular docking technology Methods The possible mechanism of ligustilide regu-lating H-type blood vessels to prevent osteoporosis was predicted by network pharmacology.Molecular docking technology was used to verify the binding ability of the core target EGFR to ligustilide.The rat model of osteo-porosis was established and divided into the sham group,model group,ligustilide high,medium and low dose(80,40,20 mg·kg-1)groups.The pathological changes of femur were observed by HE staining.The expressions of CD31,EMCN,OSX+and RUNX2+pro-tein in tibial metaphysis were detected by immunofluo-rescence.The expression of p-EGFR,p-PI3K and p-Akt protein was detected by Western blot.Results The results of network pharmacology showed that a total of 20 intersection targets were obtained.EGFR,PTGS2,ESR1 and ICAM1 were core targets,and mo-lecular docking showed that EGFR had a strong bind-ing ability with ligustilide.The signaling pathways of ligustilide in the prevention and treatment of osteoporo-sis by regulating the expression of H-type blood vessels were mainly enriched in PI3K-Akt,TNF,etc.Com-pared with the model group,ligustilide could signifi-cantly increase the number of trabecular bone and im-prove the destruction of bone microstructure.The ex-pression of CD31,EMCN,OSX+and RUNX2+signifi-cantly increased(P＜0.01,P＜0.05),the formation of H-type blood vessels were promoted,and the expres-sion of p-EGFR,p-PI3K and p-Akt significantly in-creased(P＜0.01,P＜0.05).Conclusions Ligusti-lide can increase the expression of H-type blood vessels in bone tissue of osteoporosis model rats,reduce the damage of bone trabecula and improve bone micro-structure effectively.EGFR-mediated PI3K/Akt signa-ling pathway may be the key way to exert its biological effects.

Objective To observe the value of automated fiber quantification(AFQ)for analyzing relationships of abnormal white matter microstructures and executive dysfunction in children with attention deficit hyperactivity disorder(ADHD).Methods Brain diffusion tensor imaging(DTI)and white matter structure AFQ analysis were prospectively performed on 83 children with ADHD(ADHD group)and 52 healthy children(healthy control[HC]group).The axial diffusivity(AD),radial diffusivity(RD),mean diffusivity(MD),and fractional anisotropy(FA)of complete fiber bundles and respective segments were obtained and compared between groups.Partial correlation analysis was conducted of fiber bundle diffusion parameters and the scores of Swanson Nolan and Pelham-Ⅳ(SNAP-Ⅳ)and behavior rating inventory of executive function(BRIEF)in ADHD group.Receiver operating characteristic curves were applied to assess the efficacy of each diffusion parameter for evaluating ADHD.Results AD and MD of right anterior thalamic radiation(ATR),and AD of right cingulum bundle(CB)in ADHD group were all higher than those in HC group(FWE corrected,all P＜0.000 7).Significant differences of AD of the 54-78 segments of right ATR,MD of the 13-24 and 53-77 segments of right ATR,AD of the 24-26,53-56 and 93-97 segments of right CB,as well as MD of the 72-85 segments of left inferior longitudinal fasciculus(ILF)were found between groups(FWE corrected,all P＜0.001).In ADHD group,AD of 54-78 segments of right ATR were negatively correlated with the attention deficit scores,while of 24-26 segments of right CB were negatively correlated with the emotional control scores.MD of right ATR and some segments,AD of right CB and some segments in ADHD group were positively correlated with the scores of task initiation,planning,organization,monitoring and metacognitive index(all P＜0.05).The area under the curve(AUC)of segmental fiber bundle diffusion parameter and intact fiber bundle diffusion parameter for evaluating ADHD was 0.881 and 0.808,respectively,the former was higher than the latter(Z=2.435,P＜0.05).Conclusion AFQ could be used to analyze relationships of abnormal white matter microstructures and executive dysfunction in ADHD,which could be effectively evaluated based on specific fiber bundle segments.