To understand the background of Escherichia coli(E.coli)carried by artificially bred sika deer and the biological characteristics of the isolated strains,such as drug resistance and pathogenicity,in April 2024,we collected 184 fresh deer fecal samples from four deer farms in Luxiang Township,Shuangyang District,Changchun City,Jilin Province,for isolation and cultivation of E.coli.The isolates were tested for drug resistance and biochemical identification with a BD PhoenixTM-100 Automated Microbiology System.The virulence genes were detected with PCR,and the strains were molecularly typed with ERIC-PCR.A total of 165 E.coli strains were isolated from 184 samples of deer feces,with an isolation rate of 89.67%.Twenty strains had a drug resistance phenotype,and the drug resistance rate was 12.12%;these strains included 15 strains of multi-drug resistant bacteria and 11 strains of ESBL-producing bacteria.Virulence gene detection indicated that the sika deer isolates carried multiple diarrhea-associated virulence genes,such as EAST-1(12.12%),eae(1.21%),stx1(7.88%),stx2(7.27%),and STa(1.82%).ERIC-PCR demonstrated that the isolates showed high polymorphism.The ESBL-producing E.coli carried by sika deer are likely to spread drug resistance in the community and livestock population.Some isolates carried multiple diarrhea-associated virulence genes,thus posing a human transmission risk.Therefore,monitoring of drug resistance and virulence genes must be strengthened,and antibiotics must be used reasonably during the breeding process to avoid excessive use and misuse.

Aim To explore the mechanisms of PM2.5 exposure exacerbating bleomycin(BLM)-induced idio-pathic pulmonary fibrosis(IFP)by regulating ferropto-sis via nuclear factor 2 related factor 2(Nrf2)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase(GPX)4 axis.Methods Forty C57BL/6J mice were randomized into the control,BLM,PM2.5,BLM+PM2.5 and sulforaphane(SFN,Nrf2 agonist)groups,with eight mice in each group.PM2.5 expo-sures were conducted to the BLM-induced IPF mice for two weeks.The lung function was measured,and the content of hydroxyproline(HYP)in lung tissue and the pathomorphology of lungs were observed.Reactive oxygen species(ROS),malondialdehyde(MDA),ferrous ion(Fe2+)and glutathione(GSH)of the lung tissue were measured by ELISA.The mRNA and pro-teins levels of Nrf2,SLC7A11,GPX4,collagen typeⅠ(COL-1),α-smooth muscle actin(α-SMA)were measured by quantitative polymerase chain reaction(qPCR)and Western blot.Results Compared with the control group,the lung function of mice was signif-icantly reduced(P＜0.01)in the BLM and PM2.5 groups,while lung tissue showed the characteristic pathological changes of pulmonary fibrosis such as a large number of inflammatory cell infiltration,alveolar wall fracture,thickening,collagen deposition,and sig-nificantly increased HYP,Fe2+,ROS,MDA(P＜0.05,P＜0.01),genes and proteins of COL-1,α-SMA(P＜0.01);and decreased GSH,Nrf2,SLC7A11,GPX4 genes and proteins(P＜0.05,P＜0.01).The above-mentioned lesions were markedly aggravated in the BLM+PM2.5 group compared with the BLM(P＜0.05)and PM2.5 groups(P＜0.01),and were also improved in the SFN group(P＜0.05,P＜0.01).Conclusions PM2.5 exposures can exac-erbate IPF-induced IPF in mice,and the regulating of Nrf2/SLC7 A1 1/GPX4 axis and ferroptosis might be in-volved in the related mechanisms.

Objective:To explore the value of serum P-selectin,Interleukin-18(IL-18),and Homocysteine(Hcy)levels in predicting the adverse outcomes of interventional therapy in elderly patients with deep vein thrombosis(DVT)in the lower extremities.Methods:A total of 165 elderly patients with DVT from January 2020 to August 2023 were se-lected.All patients underwent transcatheter intervention thrombolysis therapy.Patients were divided into good outcome group and poor outcome group according to the clinical treatment outcome.Serum P-selectin,IL-18,and Hcy levels were compared between the two groups.The correlation between serum P-selectin,IL-18,and Hcy levels and disease indicators,as well as the outcomes of interventional therapy was analyzed.The predictive value of serum P-selectin,IL-18,and Hcy levels for the outcomes of interventional therapy was evaluated.Results:The total effective rate of catheter-based interventional thrombolysis in 165 patients was 81.82%(135/165).The patients with good outcomes were included in the good outcome group,and the patients with poor outcomes were included in the poor outcome group.The difference in circumference between the affected side and the healthy side of the thigh and the difference in circumference between the affected side and the healthy side of the calf in the poor outcome group were greater than those in the good outcome group(P<0.05).The levels of serum P-selectin,IL-18,and Hcy in the poor outcome group were higher than those in the good outcome group(P<0.05).The levels of serum P-selectin,IL-18,and Hcy were posi-tively correlated with the difference in circumference between the affected and healthy thighs and the difference in cir-cumference between the affected and healthy calves,and negatively correlated with the outcome of interventional therapy(P<0.05).Before and after correcting other factors,serum P-selectin,IL-18,and Hcy were all independent risk factors for the outcome of interventional therapy in elderly patients with DVT(P<0.05).The area under the curve(AUC)of serum P-selectin,IL-18,and Hcy for predicting the outcome of interventional therapy for elderly DVT was 0.789(95%CI:0.718-0.848),0.812(95%CI:0.744-0.868),and 0.792(95%CI:0.722-0.851),respectively.The cutoff values were 44.21 ng/mL,185.73 ng/L,and 23.60 μmol/L,with sensitivities of 70.00%,70.00%,and 80.00%,and specificities of 82.22%,82.96%,and 68.15%,respectively.The AUC of the combined prediction of the outcomes of interventional therapy for elderly DVT was 0.935(95%CI:0.886-0.967),with a cutoff value of 44.08 ng/mL,185.16 ng/L,and 23.37 μmol/L for the three variables,with a sensitivity of 86.67%and a specificity of 88.15%.This was significantly better than the individual prediction values of the three variables(Z=5.817,4.753,5.206,all P<0.001).Conclusion:Serum P-selectin,IL-18,and Hcy are significantly correlated with disease indicators and interventional treatment outcomes in el-derly patients with DVT,and can effectively predict the outcome of interventional treatment,with a high combined pre-dictive value.

AIM To study the chemical constituents from Dictamni Cortex.METHODS The 70％ethanol extract from Dictamni Cortex was isolated and purified by HP-20 macroporous resin,silica gel,MCI,ODS and preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Thirty-three compounds were isolated and identified as rutin(1),apigenin(2),catechin(3),hesperetin(4),leonuriside A(5),androsin(6),2-methoxy-4-acetylphenol-O-α-rhamnopyranosyl-(1"-6')-β-glucopyranoside(7),vanillic acid(8),gallic acid(9),4-hydroxybenzoic acid(10),benzoic acid(11),involcranoside B(12),benzyl β-D-glucopyranoside(13),bphenylethyl-rutinoside(14),1-bromonaphthalene(15),cimifugin(16),9(S),12(S),13(S)-trihydroxyoctadeca-10(E),15(Z)-dienoic acid(17),methyl-9,12,13-trihydroxyoctadeca-10,15-dienoate(18),7,8-dihydroxy-9,12(Z,Z)-octadecadienoic acid(19),vernolic acid(20),9,10(erythro)-dihydroxy-11 E-octadecadienoic acid methyl ester(21),(7Z,9E,13Z)-11-hydroxyhexadeca-7,9,13-trienoic acid(22),(7Z,10Z,14E,16Z,19Z)-13-hydroxydocosa-7,10,14,16,19-pentaenoic acid(23),(9E)-8,11,12-trihydroxyoctadecenoic acid methyl ester(24),n-hexanol-O-rutinoside(25),hexyl β-sophoroside(26),3-pentyl 6'-(3-hydroxy-3-methylglutaryl)-β-D-glucopyranoside(27),3-methylbut-3-enyl-6-O-β-D-glucopyranosyl-β-D-glucopyranoside(28),3-methyl-but-2-en-1-yl β-D-glucopyranoside(29),3-methylbutan-1-ol-β-D-glucopyranoside(30),pregnenolone(31),2-butoxytetrahydrofuran(32),psydrin(33).CONCLUSION Compounds 2-4,8-13,15-16,25-28 and 32-33 are isolated from Rutaceae family for the first time.

Immobilized enzyme-based enzyme electrode biosensors, characterized by high sensitivity and efficiency, strong specificity, and compact size, demonstrate broad application prospects in life science research, disease diagnosis and monitoring, etc. Immobilization of enzyme is a critical step in determining the performance (stability, sensitivity, and reproducibility) of the biosensors. Random immobilization (physical adsorption, covalent cross-linking, etc.) can easily bring about problems, such as decreased enzyme activity and relatively unstable immobilization. Whereas, directional immobilization utilizing amino acid residue mutation, affinity peptide fusion, or nucleotide-specific binding to restrict the orientation of the enzymes provides new possibilities to solve the problems caused by random immobilization. In this paper, the principles, advantages and disadvantages and the application progress of enzyme electrode biosensors of different directional immobilization strategies for enzyme molecular sensing elements by specific amino acids (lysine, histidine, cysteine, unnatural amino acid) with functional groups introduced based on site-specific mutation, affinity peptides (gold binding peptides, carbon binding peptides, carbohydrate binding domains) fused through genetic engineering, and specific binding between nucleotides and target enzymes (proteins) were reviewed, and the application fields, advantages and limitations of various immobilized enzyme interface characterization techniques were discussed, hoping to provide theoretical and technical guidance for the creation of high-performance enzyme sensing elements and the manufacture of enzyme electrode sensors.

Objective: To investigate the effect of neurite outgrowth inhibitor extracellular peptide residues 1-40 (NEP1-40) combined with poly (lactic-co-glycolic acid) (PLGA) and gelatin electrospun fiber membrane on facial nerve repair in rats. Methods: According to the principle of random grouping, 108 male SD rats were divided into four groups (n = 27 in each group, approved by the ethics committee), namely, the sham group, control group, PLGA group, and NEP1-40 + PLGA group. A facial nerve fracture model was established for all of the groups except for the sham group. The control group received no further treatment, the PLGA group and the NEP1-40+PLGA group were supported by PLGA membrane, and the NEP1-40+PLGA group received one immediate local injection of NEP1-40 (5 μg/μL) at a dose of 10 μL. Facial nerve function analysis, electrophysiological examination, transmission electron microscope observation, HE staining, and immunohistochemical staining of myelin marker S100β and axonal marker β3-tubulin were used to evaluate the recovery of injured facial nerves of rats at 2, 4 and 8 weeks. Results : At 8 weeks, the facial nerve function score of the NEP1-40+PLGA group was better than that of the control group and PLGA group (P < 0.001), and facial nerve function was significantly restored. Electrophysiological examination of nerve action potentials at the injured facial nerve showed that the amplitude in the NEP1-40+PLGA group was higher than that of the control group and PLGA group (P < 0.001), but there was no significant difference in latency and conduction velocity results between the groups (P > 0.05). At 2, 4, and 8 weeks, transmission electron microscopy showed that the number of myelinated nerve fibers and myelin sheath thickness in the cross-section of the injured facial nerve in the NEP1-40+PLGA group were greater than those in the other groups (P < 0.05). At 8 weeks, HE staining showed that the facial nerves in the control group had partially recovered, but the overall cell distribution was uneven and the boundary with surrounding tissues was slightly blurred. In contrast, the NEP1-40+PLGA group had a relatively uniform cell distribution and a clearer boundary with surrounding tissues. At 2, 4, and 8 weeks, the immunohistochemical results showed that in the cross-section of the injuried facial nerve, NEP1-40 increased the expression of neural markers S100 β and β3-tubulin, especially β3-tubulin, which was close to normal levels (P > 0.05) Conclusion NEP1-40 is beneficial for the generation of new myelin sheaths and axons at the site of injury, and it can promote the repair and regeneration of injured facial nerves to a certain extent, thus accelerating the recovery of injured nerve function.

Objective To investigate the neuroprotective effects and mechanisms of abscisic acid(ABA)in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced Parkinson's disease(PD)mouse models.Methods Eight-week-old C57BL/6J mice were randomly divided into three groups,control group(Ctrl),MPTP group,and MPTP+ABA group,12 mice in each group.Except for the control group,mice in the other groups were intraperitoneally injected with MPTP 25 mg/kg daily for 8 consecutive days to establish a subacute PD model.The MPTP+ABA group received intraperitoneal injections of ABA 25 mg/kg daily for 11 consecutive days,starting 3 days prior to MPTP administration.Behavioral tests were performed 24 hours after the last administration.On day 3,the expression of tyrosine hydroxylase(TH)and glial fibrillary acidic protein(GFAP)in the substantia nigra pars compacta(SNc)and striatum(STR)was analyzed by Western blotting,and mRNA levels of inflammatory factors were measured by Real-time PCR.Immunofluorescent staining was used to detect the expression of TH,GFAP,and ionized calcium-binding adapter molecule 1(Iba1).Results Compared with the control group,MPTP-treated mice exhibited impaired motor function,a reduced number of TH-positive dopaminergic neurons in the SNc,down-regulated TH protein expression in both the SNc and striatum,up-regulated GFAP protein expression,increased numbers of GFAP-and Iba1-positive cells,and elevated levels of pro-inflammatory factors.In contrast,the MPTP+ABA group showed improved motor function,increased TH-positive neurons in the SNc,up-regulated TH protein expression,down-regulated GFAP protein expression,reduced numbers of GFAP-and Iba1-positive cells,and decreased pro-inflammatory factor levels compared to the MPTP group.Conclusion ABA ameliorates motor dysfunction in MPTP-induced PD model mice,reduces degeneration and death of dopaminergic neurons in the SNc,suppresses the proliferation and activation of astrocytes and microglia in the SNc and striatum,and alleviates neuroinflammation.These results suggest that ABA exerts neuroprotective effects in MPTP-induced PD model mice.

Objective To find out whether photobiomodulation(PBM)can mitigate cognitive dysfunction caused by chronic stress by affecting levels of adenosine triphosphate(ATP)and adenosine receptors.Methods Twenty-four C57BL/6J mice were randomly divided into a control group,a stress group,and a treatment group.Chronic unpredictable mild stress was used to establish a mouse model of stress.Six weeks into modeling,the treatment group was subjected to one week of PBM interventions.Behavioral tests were conducted to observe behavioral changes in the mice.Western blotting(WB)was used to detect the expressions of A1,A2B,and A3 adenosine receptors in the hippocampus and prefrontal cortex of mice in the three groups.Twelve C57BL/6J mice were randomly divided into a control group and an intervention group.The intervention group received a week of PBM interventions and underwent behavioral testing.WB was used to detect the expression changes of A1,A2B,and A3 adenosine receptors in the hippocampus and prefrontal cortex in both groups.Immunofluorescence assay was adopted to detect the expression of c-Fos in the hippocampus of mice in the two groups.The ATP assay kit made by Beyotime Biotechnology Co.,Ltd.was used to measure changes in ATP contents in the hippocampus and prefrontal cortex tissues of mice.Cell experiments were conducted to verify the effect of PBM on intracellular ATP contents.Results Mice in the stress group covered a similar distance to the control group,but finished far fewer platform crossings.There was no significant difference between the treatment group and the control group in the number of times of platform crossings,but compared favorably with the stress group where the levels of adenosine receptors in the hippocampus and prefrontal cortex were lower,but were increased by PBM.After PBM interventions in normal mice,platform crossings were increased significantly compared to the control group.PBM also raised adenosine receptor levels and ATP contents in the hippocampus and prefrontal cortex,and increased hippocampal c-Fos expressions.In vitro,PBM elevated intracellular ATP levels.Conclusion PBM may improve chronic stress-induced cognitive dysfunction by regulating ATP levels and adenosine receptor expressions,thereby modulating neuronal responsiveness in the hippocampus.

Objective:To explore the effect of repetitive transcranial magnetic stimulation (rTMS) on the cognition and ability in the activities of daily living (ADL) of persons with cerebral small vessel disease.Methods:A total of 71 patients with cognitive impairment due to cerebral small vessel disease were randomly divided into a control group ( n=35) and a treatment group ( n=36). The treatment group was further divided into a group with mild cognitive impairment (a score of 21 to 26 on the mini-mental state exam, n=23) and a moderate cognitive impairment group (scoring 10 to 20, n=13). The treatment groups received 10Hz rTMS, while the control group received sham rTMS. Before and after 90 days of treatment, all were assessed using the mini-mental state examination (MMSE) and the cognitive subscale of the vascular dementia assessment scale (VaDAS-cog). Ability in the activities of daily living was quantified using the Barthel Index (BI). Results:After treatment, significantly increased MMSE and BI scores, but significantly decreased VaDAS-cog scores were observed in both groups. At that point the treatment group showed a significantly higher average post-treatment MMSE score (25.61±2.94) and BI score (80.00±7.85), but a significantly lower average VaDAS-cog score (44.06±3.41) than the control group. The average MMSE scores were not significantly different between the mild and moderate cognitive impairment subgroups. No severe adverse reactions were observed during the rTMS treatment.Conclusions:The 10Hz rTMS can effectively improve the cognition and daily living ability of persons with cerebral small vessel disease.

Objective"Lie flat"has recently become a buzz word describing a specific psychological state in some individuals.However,its psychological meaning and mechanisms remain unclear.Lack of motivation under certain conditions may be one of the key psychological characteristics of this condition.Methods Sixteen male SD rats were randomly assigned to two food-restriction levels(90%and 80%of baseline weight,respectively),and subjected to a sucrose-pellet self-administration task.The establishment of the model is divided into three stages.The rats were trained to self-administer sucrose pellets in a high-reward learning stage and in a low-reward learning stage.We then assessed"lie flat"behavior in a preference-test stage,in which rats could choose between high-reward-and low-reward-paired nose-pokes in a discrete choice procedure.Results Combining self-feeding training with Western blot,the results showed that:(1)rats showed significantly decreased nose-poke behavior for low-reward,i.e."lie flat"behavior,but not for high-reward,when≥90%of the maximal reward was obtained in the high-reward task and the effort(X)in the low-reward task increased to the breaking point.(2)Dopamine D1R expression in the NAc was significantly higher in the"lie flat"group compared with the"not lie flat"rats,while D2R expression was similar in both groups.Conclusions Rats can show"lie flat"behavior,and up-regulation of D1R expression in the NAc may be a key part of the molecular basis responsible for this motivation deficiency in"lie flat"behavior.These results extend our understanding of"lie flat"behavior,and provide a new paradigm for the study of its mechanism.