ObjectiveTo investigate the effects of hypoxia-treated mesenchymal stem cell （MSCs） exosomes （Exo） and their loading with curcumin on microglial inflammatory responses， and to explore the enhancing effect of hypoxia treatment on the function of MSCs Exo. MethodsThe supernatants of human umbilical cord （hUC）-MSCs cultured under normal and hypoxic conditions were collected， and Exo were isolated using ultracentrifugation. After identification by transmission electron microscopy and Western blot， curcumin was loaded using the co-incubation method. The lipopolysaccharide （LPS）-induced microglial inflammation model was treated with dimethyl sulfoxide （DMSO）， curcumin， normoxia Exo， hypoxia Exo， normoxic Exo loaded with curcumin， and hypoxic Exo loaded with curcumin， respectively. The expression of the M1-type marker inducible nitric oxide synthase （iNOS） in BV2 cells was detected by immunofluorescence （IF）. Western blot and enzyme-linked immunosorbent assay （ELISA） were used to measure the expression and secretion levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and IL-6 in the cells and their culture supernatants. ResultsNormoxia Exo， hypoxia Exo， normoxic Exo loaded with curcumin， and hypoxic Exo loaded with curcumin exhibited a "saucer-like" shape with a diameter ranging from 30~150 nm， and the expression of exosomal markers CD9， CD81， and TSG101 were positive. After treating the BV2 cell inflammation model， IF results showed that， compared with the normoxia Exo group， treatment with hypoxic Exo significantly reduced the expression of iNOS. Moreover， when compared with the curcumin group and the normoxic Exo loaded with curcumin group， the expression level of iNOS significantly decreased after treatment with hypoxic Exo loaded with curcumin. The results of Western blot and ELISA indicated that， in comparison with the normoxia Exo group， treatment with hypoxic Exo significantly reduced the expression and secretion of the inflammatory cytokines TNF-α， IL-1β， and IL-6. Additionally， when compared with the curcumin group and the normoxic Exo loaded with curcumin group， both the expression and secretion of TNF-α， IL-1β， and IL-6 significantly decreased after treatment with hypoxic Exo loaded with curcumin. ConclusionHypoxia preconditioning can enhance the ability of hUC-MSCs-Exo in the inhibition of microglial polarization and inflammatory factors’ secretion. Additionally， using Hypoxia-MSCs-Exo as a drug-delivery carrier of curcumin can improve its solubility and stability， facilitating its absorption by cells and exerting the therapeutic effect of combination therapy.

Objective: To analyze the causes of abnormal blood agglutination caused by high-titer cold antibodies in blood donors from a serological perspective. Methods: The donor's blood type was identified using the tube method. Direct antiglobulin test (DAT), indirect antiglobulin test (IAT), and antibody titer detection were performed on the samples. Results: The blood type of the donor was type B, and the DAT was negative. No complement components were detected on the surface of the donor's red blood cells. The supernatant of the suspended red blood cells was clear without abnormal coloration, and the agglutinated clumps were intact and difficult to dissociate. The antibody was identified as an IgM-type anti-IH cold agglutinin. At 4℃, this antibody reacted with autologous red blood cells, adult group B red blood cells, adult group O red blood cells, cord blood group B red blood cells and cord blood group O red blood cells, with a stronger reaction intensity observed with group O cells than with B cells. The titers were 512, 128, 256, 32 and 128, respectively. Conclusion: High-titer anti-IH can cause abnormal agglutination of blood at low temperatures. During blood distribution and clinical use, the appearance of blood products shall be strictly inspected to prevent the release of non-conforming blood products.

ObjectiveTo investigate the mechanisms by which the combination of berberine （BBR） and baicalin （BAI） ameliorates type 2 diabetes mellitus （T2DM） due to internal accumulation of dampness-heat from the perspectives of gut microbiota and metabolomics. MethodsAntibiotics were used to induce pseudo-sterile mice. Thirty pseudo-sterile mice were randomized into a normal fecal microbiota transplantation group （n=10） and a T2DM （syndrome of internal accumulation of dampness-heat） fecal microbiota transplantation group （n=20）. The mice were then administrated with suspensions of fecal microbiota from healthy volunteers and a patient with T2DM due to internal accumulation of dampness-heat by gavage， respectively. Each mouse received 200 µL suspension every other day for a total of 15 times to reshape the gut microbiota. The T2DM model mice were then assigned into a model group （n=8） and a BBR-BAI group （n=11）. BBR was administrated at a dose of 200 mg·kg^-1， and BAI was administrated in a ratio of BBR-BAI 10∶1 based on preliminary research findings. The administration lasted for 8 consecutive weeks. Fasting blood glucose （FBG）， glycated hemoglobin （HbA1c）， insulin （INS）， triglycerides （TG）， total cholesterol （CHOL）， low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C）， aspartate aminotransferase （AST）， and alanine aminotransferase （ALT） levels were measured to evaluate the effects of the BBR-BAI combination on glucose and lipid metabolism and liver function in T2DM mice. Hematoxylin-eosin staining was employed to observe pathological changes in the colon tissue. The expression of claudin-1， zonula occludens-1 （ZO-1）， and occludin in the colon tissue was determined by Western blot. Real-time quantitative polymerase chain reaction（Real-time PCR） was employed to assess the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） in the colon tissue. The fecal microbiota composition and differential metabolites were analyzed by 16S rRNA sequencing and ultra-high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry （UPLC-Q-TOF-MS）， respectively. ResultsThe BBR-BAI combination lowered the FBG， HbA1c， and INS levels （P<0.05， P<0.01） and alleviated insulin resistance （P<0.01） in T2DM mice. Additionally， BBR-BAI elevated the levels of ZO-1， occludin， and claudin-1 （P<0.05， P<0.01） and down-regulated the expression levels of TNF-α， IL-1β， and IL-6 in the colon （P<0.05， P<0.01）. The results of 16S rRNA sequencing showed that BBR-BAI increased the relative abundance of Ligilactobacillus， Phascolarctobacterium， and Akkermansia （P<0.05）， while significantly decreasing the relative abundance of Alistipes， Odoribacter， and Colidextribacter （P<0.05）. UPLC-Q-TOF-MS identified 28 differential metabolites， which were primarily involved in arachidonic acid metabolism and α-linolenic acid metabolism. ConclusionBBR-BAI can ameliorate T2DM due to internal accumulation of dampness-heat by modulating the relative abundance of various bacterial genera in the gut microbiota and the expression of fecal metabolites.

AIM: To evaluate the effectiveness and safety of compound anisodine combined with laser photocoagulation in the treatment of diabetic retinopathy(DR).METHODS: A prospective cohort study was used to select 80 patients(160 eyes)diagnosed with severe non-proliferative diabetic retinopathy(NPDR)and proliferative diabetic retinopathy(PDR)in Beijing Tongren Hospital, Capital MedTcal University and Beijing Daxing District People's Hospital from May 2023 to July 2023. They were divided into control group(40 cases, 80 eyes)and observation group(40 cases, 80 eyes)by random number table method. The control group only received 532 nm laser panretinal photocoagulation(PRP)treatment, while the observation group received PRP treatment together with superficial temporal subcutaneous injection of compound anisodine. The clinical efficacy, changes in hemorheology, changes in retinal blood vessels, and incidence of adverse reactions in the two groups were observed before and at 2 mo after treatment.RESULTS: The visual acuity, fundus changes and hemorheological parameters of the two groups were analyzed before and after treatment. There were no significant differences in the two groups before treatment(all P>0.05). The best corrected visual acuity of the observation group was better than that of the control group at 2 mo after treatment(P<0.05), and the clinical curative effect of fundus was also better than that of the control group(all P<0.05). The hemorheological indexes of central retinal artery blood flow(peak systolic velocity and end diastolic velocity)in the observation group were higher than those of the control group(all P<0.05), and the blood flow resistance index was lower than that of the control group(P<0.05).CONCLUSION: Compound anisodine combined with 532 nm laser photocoagulation is safe and effective in the treatment of DR, and the visual recovery effect is better.

Objective:To analyze the relationship between neutrophil-to-lymphocyte ratio(NLR) and the levels of inflammatory factors in peripheral blood and the brain injuries of suppurative meningitis in neonates (NPM) and the value of joint prediction of hydrocephalus.Methods:Ninety-five children with NPM diagnosed and treated in the Affiliated Hospital of Jining Medical College from January 2020 to December 2022 were retrospectively selected, including 31 children with non-cerebral parenchymal injury (group A), accounting for 32.63%(31/95); 36 children with parenchymal injury and no hydrocephalus (group B), accounting for 37.89%(36/95); 28 children with brain parenchymal injury and hydrocephalus (group C), accounting for 29.47%(28/95). The clinical data, peripheral blood levels of NLR, interleukin 6(IL-6), hyper-sensitive C-reactive protein (hs-CRP) and procalcitonin (PCT) were compared among the three groups, and the influencing factors of hydrocephalus were analyzed by Logistic regression. Receiver operating characteristic (ROC) curve was used to analyze the value of changes(△, representing the absolute value of the difference between the corresponding indexes and 3 d after admission) in NLR, hs-CRP, IL-6 and PCT in predicting hydrocephalus in NPM alone and combination.Results:The incidence of respiratory failure and convulsion and the levels of NLR, hs-CRP, IL-6 and PCT in the group C were higher than those in the group B and A: 42.86%(12/28) vs. 13.89%(5/36), 6.45%(2/31); 42.86%(12/28) vs. 16.67%(6/36), 3.23%(1/31); 3.91 ± 0.71 vs. 3.12 ± 0.64, 2.16 ± 0.58; (98.21 ± 10.17) mg/L vs. (86.74 ± 9.30), (75.18 ± 8.42) mg/L; (40.73 ± 7.88) ng/L vs. (31.26 ± 6.45), (25.01 ± 5.19) ng/L; (13.26 ± 2.04) ng/L vs (11.83 ± 1.91), (9.45 ± 1.67) ng/L; the levels of NLR, hs-CRP, IL-6 and PCT in the group B were higher than those in the group A, there were statistical differences ( P<0.05). Logistic regression analysis showed that the levels of peripheral blood NLR and serum hs-CRP, IL-6 and PCT at admission were independent risk factors for hydrocephalus in children with NPM ( P<0.05). The △NLR, △hs-CRP, △IL-6 and △PCT of the children with hydrocephalus were lower than those without hydrocephalus: 0.73 ± 0.24 vs. 1.02 ± 0.31, (42.73 ± 13.08) mg/L vs. (69.56 ± 18.19) mg/L, (13.43 ± 4.22) ng/L vs. (19.86 ± 6.43) ng/L, (4.35 ± 1.34) ng/L vs. (6.58 ± 2.03) ng/L, there were statistical differences ( P<0.05). ROC curve analysis results showed that the AUC of △NLR, △hs-CRP, △IL-6 and △PCT combined to predict hydrocephalus in children with NPM was 0.903, which was greater than each single index ( P<0.05). Conclusions:The levels of NLR, hs-CRP, IL-6 and PCT in peripheral blood of children with NPM are significantly correlated with brain injuries, and have certain predictive efficacy in predicting the risk of hydrocephalus.

Objective To explore the application effect of bedside critical ultrasound in volume management of acute kidney injury(AKI)donors.Methods Clinical data of 56 AKI donors and 106 recipients from the Transplantation Center of People's Hospital of Guangxi Zhuang Autonomous Region from October 1,2020 to May 31,2022 were collected.They were divided into the critical ultrasound group(34 donors,66 recipients)and the traditional central venous pressure(CVP)group(22 donors,40 recipients)according to the volume management methods.The AKI stage and recovery time,renal function indicators(serum creatinine(Scr),cystatin C,estimated glomerular filtration rate),donor Remuzzi score,cold ischemia time,biventricular inotrope usage rate and time,delayed graft function(DGF)incidence and recovery time,and renal function indicators at 6 and 12 months after surgery were compared and analyzed between the two groups.Results There were no statistically significant differences in gender,age,body weight,AKI stage,pre-acquisition renal function indicators,biventricular inotrope usage rate,renal function indicators at 6 and 12 months after surgery,DGF recovery time,donor Remuzzi score and cold ischemia time of the donors and recipients between two groups(all P>0.05).The AKI recovery time,continuous renal replacement therapy rate and biventricular inotrope usage time of donors in the critical ultrasound group were shorter or lower than those in the traditional CVP group(all P<0.05).The incidence of DGF in recipients of the critical ultrasound group was lower than that of the traditional CVP group(P<0.05).Subgroup analysis showed that there was no statistically significant difference in Scr at 6 and 12 months after surgery in recipients of the critical ultrasound group(P>0.05),while the Scr at 12 months after surgery was higher than that at 6 months in recipients of the traditional CVP group(P<0.05).Conclusions AKI kidneys may be used for kidney transplantation after active maintenance.Bedside critical ultrasound has unique advantages in volume management of AKI donors and may improve the function of AKI kidneys to a certain extent.

Objective To explore differentially expressed proteins in the serum exosomes of acute myeloid leukemia(AML)patients and healthy controls by using the proteomics method,and provide new biomarker for the diagnosis and treatment of AML.Methods A total of 76 AML patients diagnosed and treated in the First People's Hospital of Honghe State from November 2022 to June 2024 were selected as the experimental group,and 60 healthy physical examination participants were selected as the control group.Tandem Mass Tag(TMT)labeled proteomics was used to identify the differences in protein expression in serum exosomes between the experimental and control groups,and the identified differentially expressed proteins were subjected to bioinformatics analysis.Western-Blot was used to verify the differentially expressed proteins.Results Comparedwith healthy controls,146 aberrantly expressed proteinswere detected in serum exosomes of AML patients,of which 89 were up-regulated and 57 were down-regulated.Among them,Alpha-1-antichymotrypsin(α1-ACT),Angiogenin(ANG),Vitronectin(VIT),Clusterin(Clu),Fibronectin(FN),Keratin type I cytoskeletal-18(KRT18),and actinin alpha4(ACTN4)showed changed significantly.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis indicated that differentially expresseproteins were involved in biological processes such as neutrophil immunity,neutrophil degranulation,neutrophil activation immune response,and protein activation cascade,significantly enriched in 25 biological pathways including complement and coagulation cascades,extracellular matrix(ECM)-receptor interaction,and hypoxia-inducible factor-1(HIF-1)signaling pathways.Western-blot validation results showed that compared with the control group,the expression levels of ANG,Clu and FN in the serum exosomes of AML patients were significantly increased,while the expression level of ACTN4 was significantly decreased,with statistically significant differences(t=-11.854～18.569,all P<0.05).The expression levels of ANG and FN were correlated with white blood cell count(F=8.888,7.818,all P<0.05)and cytogenetics(F=8.619,7.983,P<0.05).The expression level of Clu protein was correlated with white blood cell count(F=2.571,P<0.05),but not significantly associated with cytogenetics(F=1.886,P>0.05).ANG,FN and Clu showed no significant correlation with FAB classification(F=0.175,0.434,0.042,all P>0.05).Conclusion The expression of exosomal proteins in the serum of AML patients compared to healthy individuals are significant differences,which may serve as serological markers for the diagnosis and treatment of AML.

Objective To explore the biological functions and mechanisms of PIWI-interacting RNA(piRNA)in cervical cancer(CC).Methods RT-qPCR was used to detect the expression level of piRNA-2732 in CC tissue,CaSki cells and End1/E6E7 cells.EpiQuik m6A RNA methylation quantification kit was used to detect the methylation level of m6A RNA in CaSki cells.The expression levels of methyltransferases(METTL3,METTL14 and WTAP)and demethylases(FTO,ALKBH5)mRNA in CaSki cells were detected by RT-qPCR.After culturing CaSki cells to logarithmic growth stage,they were divided into six groups:piRNA-2732 mimic negative control group(mi-NC group),piRNA-2732 mimic group(mi-2732 group),piRNA-2732 inhibitor negative control group(in-NC group),piRNA-2732 inhibitor group(in-2732 group),piRNA-2732 mimic+METTL3 knockdown control group(mi-2732+si-NC group),and piRNA-2732 mimic+METTL3 knockdown group(mi-2732+si-METTL3 group).The viability of CaSki cells was detected by CCK8 assay.Colony formation assay was used to detect the proliferation ability of CaSki cells.Transwell experiment was used to detect the migration and invasion ability of CaSki cells.RT-qPCR and Western blot were used to detect the expression of methyltransferase like protein 3(METTL3).Transfected METTL3 wild-type(METTL3-WT)and METTL3 mutant(METTL3-MUT)in the mi-NC group,mi-2732 group,in-NC group,and in-2732 group respectively,and detected the effect of piRNA-2732 on METTL3 through dual luciferase reporter gene assay.Results Compared with the adjacent tissues,the expression of piRNA-2732(3.84±1.08 vs 1.32±0.53)was significantly higher in the cancer tissues of CC patients,and the difference was statistically significant(t=5.115,P<0.001).Compared with end1/E6E7 cells,the expression of piRNA-2732(1.00±0.13 vs 1.67±0.16)in CaSki cells was significantly higher,and the difference was statistically significant(t=5.632,P<0.01).Compared with mi-NC group,mi-2732 group promoted the viability,proliferation,migration and invasion of CaSki cells,and the differences were statistically significant(t=4.410～11.040,all P<0.01).Compared with mi-NC group,mi-2732 group increased m6A RNA methylation level and METTL3 mRNA and protein,the differences were statistically significant(t=6.176,9.211,12.550,all P<0.05).The results of dual luciferase reporter gene testing showed that compared with the mi-NC+METTL3-WT group,the relative luciferase activity of mi-2732+METTL3-WT group was significantly increased(t=11.850).Compared with mi-2732+METTL3-WT group,the relative luciferase activity of mi-2732+METTL3-MUT group was significantly lower(t=12.740),and the difference was statistically significant(all P<0.000 1).Compared with in NC+METTL3-WT group,the relative luciferase activity of in-2732+METTL3-WT group was significantly lower(t=7.828),compared with in-2732+METTL3-WT group,the relative luciferase activity of CaSki cells in in-2732+METTL3-MUT group was significantly increased(t=8.146),and the difference was statistically significant(all P<0.001).Compared with mi-2732+si-NC group,the expression level of m6A in mi-2732+si-METTL3 group was significantly lower,and the difference was statistically significant(t=7.630,P<0.01).Compared with mi-2732+si-NC group,the proliferation ability,colony number,cell migration and invasion ability of CaSki cells in mi-2732+si-METTL3 group were significantly decreased,and the differences were statistically significant(t=3.695～4.891,all P<0.001).Conclusion piRNA-2732 is overexpressed in CC tissues and cells,and piRNA-2732 promotes tumor development in CC through METTL3 mediated m6A methylation.

Objective To evaluate answers to typical clinical questions related to neuro-ophthalmology generated by Artificial Intelligence(AI)Large Language Models(LLM)and to explore the performance of neuro-ophthalmology-related questions on LLM in a multidimensional manner using objective and expert assessment.Methods Multicenter,random-ized,cross-sectional pilot study.Thirty typical questions related to neuro-ophthalmology were selected based on four per-spectives:definition,etiology,clinical manifestations and signs,and treatment and prognosis,and were analyzed quantita-tively using Deepseek,Wenxin Yiyin 4.0,Doubao,and Kimi 1.5,which are four open-source LLMs in China,and quantita-tively analyzed with objective assessment;and quantitatively rated by three ophthalmologists using expert assessment for 120 answer texts.Three ophthalmology experts quantitatively scored the 120 answer texts.Three ophthalmologists quantita-tively scored the 120 answer texts.Level 3,5,and 4 Likert scales were developed according to the completeness,accura-cy,professionalism,relevance,and criticality of the question texts,respectively.The best-performing LLM was selected,and its performance was observed across the four types of questions.Additionally,three other experts assessed whether the best-performing one could be evaluated as a substitute for real-world doctor-patient communication.Results In the objective Chinese text reading difficulty analysis,the differences in total word count among the four LLMs were statistically significant(all P＜0.001).Of the four LLMs,Kimi 1.5 performed the best,with frequencies of 61％,29％,and 41％for the highest scores in completeness(3),accuracy and professionalism(5),and relevance and usefulness(4),respective-ly.Kimi 1.5 performed more consistently on the questions on the four areas of neuro-ophthalmologic disorders:definition,etiology,clinical manifestations and signs,treatment,and prognosis,with no between-group differences(P＞0.05).Con-clusion Chinese language LLMs have great potential in the clinical application of neuro-ophthalmology.Kimi 1.5 outper-forms other LLMs in terms of completeness,accuracy,professionalism,relevance,and usefulness,but it still cannot re-place real-world doctor-patient communication.There is a need to explore new diagnostic and therapeutic model of AI+physician in the future.

Objective To investigate the pharmaceutical monitoring for children with acute kidney injury caused by voriconazole combined with cyclosporine,in order to provide methods and ideas for clinical pharmacists to conduct pharmaceutical services.Methods The clinical pharmacist assisted physicians to analyze the causes of two episodes of abnormal renal function in the children,and used the therapeutic drug monitoring to monitor the whole process of medication.Results The child's acute kidney injury was likely to be related to medication,and physicians adopted the pharmacist's recommendation for therapeutic drug monitoring and drug adjustment,and the child didn't have any renal function abnormalities again.Conclusion Clinical pharma-cists can use pharmacokinetic knowledge and therapeutic drug monitoring methods to assist physicians in formulating and optimi-zing treatment plans,and monitor the whole process of medication for special children to ensure safety medication.