ObjectiveTo investigate the mechanism by which adrenoceptor alpha 2A (Adra2a) regulates lipopolysaccharide (LPS)-induced inflammation in primary hepatocytes from lipopolysaccharide-binding protein (LBP) knockout mice (Lbp^-/-). MethodsPrimary hepatocytes from C57BL/6J and Lbp^-/- mice were isolated using a two-step perfusion method. An in vitro inflammatory model was established by LPS stimulation, and an in vivo inflammatory mouse model was established by intraperitoneal injection of LPS. The in vitro experiments were grouped as follows: Control group, LPS group, BRL+LPS group, OE-NC+LPS group, and OE-Adra2a+LPS group. The Control group served as the blank control. The LPS group involved stimulating primary hepatocytes with LPS. The BRL+LPS group involved pretreating primary hepatocytes with BRL-44408 maleate followed by LPS stimulation. The OE-NC+LPS group involved transfecting primary hepatocytes with an empty vector followed by LPS stimulation. The OE-Adra2a+LPS group involved transfecting primary hepatocytes with a lentivirus overexpressing Adra2a, followed by LPS stimulation. The in vivo experimental groups were divided into Control', LPS', BRL+LPS', OE-NC+LPS', and OE-Adra2a+LPS' groups. The Control' group served as the blank control. The LPS' group received intraperitoneal injection of LPS. The BRL+LPS' group received intraperitoneal injection of BRL-44408 maleate for pretreatment, followed by LPS injection. The OE-NC+LPS' group received intraperitoneal injection of empty vector for pretreatment, followed by LPS injection. The OE-Adra2a+LPS' group received intraperitoneal injection of a lentivirus overexpressing Adra2a for pretreatment, followed by LPS injection. Cell viability after Adra2a inhibition and overexpression was assessed via the Cell Counting Kit-8 (CCK-8) assay. RT-qPCR measured changes in gene expression levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) after Adra2a inhibition and overexpression. Western blotting was performed to detect Adra2a protein expression and phosphorylation levels of extracellular signal-regulated kinase 1/2 (ERK1/2), p38 mitogen-activated protein kinase, and c-Jun N-terminal kinase (JNK) following LPS stimulation. ResultsIn vitro experiments revealed that LPS stimulation significantly decreased Adra2a protein expression in primary hepatocytes from C57BL/6J mice compared to the Control group (P<0.05), whereas it increased in primary hepatocytes from Lbp^-/- mice (P<0.001). Compared to the LPS group, the BRL+LPS group exhibited significantly increased cell viability (P<0.01), reduced TNF-α, IL-6, and IL-1β gene transcription levels (P<0.01, P<0.001, P<0.001), and decreased phosphorylation levels of MAPK signaling pathway-related proteins ERK1/2, p38, and JNK (P<0.01, P<0.001, P<0.001). Compared with the OE-NC+LPS group, the OE-Adra2a+LPS group showed significantly decreased cell viability (P<0.001), increased gene transcription levels of TNF-α, IL-6, and IL-1β genes (P<0.001, P<0.01, P<0.001), and elevated phosphorylation levels of MAPK signaling pathway-related proteins ERK1/2, p38, and JNK (P<0.001, P<0.01, P<0.001). In vivo experiments showed that, compared with the LPS' group, the BRL+LPS' group exhibited significantly reduced phosphorylation levels of MAPK signaling pathway-related proteins ERK1/2, p38, and JNK (P<0.001, P<0.01, P<0.01). In the OE-Adra2a+LPS' group, the phosphorylation levels of ERK1/2, p38, and JNK were significantly elevated compared to the OE-NC+LPS' group (P<0.01, P<0.001, P<0.01). ConclusionLPS stimulation can cause a significant increase in Adra2a protein expression in primary hepatocytes of Lbp^-/- mice. Adra2a protein can regulate the level of LPS-induced inflammation in primary hepatocytes of Lbp^-/- mice through the MAPK signaling pathway.

ObjectiveTo investigate the anti‑pancreatic fibrosis mechanism of Klotho‑derived peptide 7 （KL7） by observing its effect on a mouse model of chronic pancreatitis （CP） induced by cerulean， and to provide a basis for clinical medication. MethodsA total of 40 male BALB/c mice were randomly divided into control group， model group， low-dose KL7 group （2 mg/kg）， and high-dose KL7 group （4 mg/kg）， with 10 mice in each group. All mice except those in the control group were given intraperitoneal injection of cerulean （50 μg/kg） 6 times a day at an interval of 1 hour， twice a week for 4 consecutive weeks to establish a model of CP. The mice in the low-dose KL7 group and the high-dose KL7 group were treated with different doses of KL7 once a day for 4 consecutive weeks. In vivo imaging was used to observe the accumulation of KL7 in the pancreas； molecular docking was used to detect the binding of KL7 to transforming growth factor-β type Ⅱ receptor （TβRⅡ）； the mice were measured in terms of body weight and pancreatic weight； HE staining was used to observe the pathological changes of pancreatic tissue； Masson staining was used to observe the degree of pancreatic fibrosis； immunohistochemical staining was used to measure the expression of α-smooth muscle actin （α-SMA） and type Ⅰ collagen （COL1A1）； Western blotting was used to measure the protein expression levels of α-SMA， TβRII， and phosphorylated small mothers against decapentaplegic homolog 2/3 （p-Smad2/3） in pancreatic tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test and the Dunnett’s-T3 test were used for further comparison between two groups. ResultsKL7 was significantly enriched in the pancreatic tissue of CP mice， and there was a strong binding activity between KL7 and TβRⅡ. Compared with the control group， the model group had significant reductions in pancreatic mass and relative pancreatic mass （P<0.000 1）， with disordered structure of pancreatic tissue， an increase in inflammatory cell infiltration， and significant increases in fibrosis degree， the positive areas of α-SMA and COL1A1 （P<0.000 1）， and the protein expression levels of α-SMA， TβRⅡ， and p-Smad2/3 （P<0.05）. Compared with the model group， the high-dose KL7 group had significant increases in pancreatic mass and relative pancreatic mass （P<0.01）， with alleviation of structural damage of pancreatic tissue and inflammatory cell infiltration， a significant reduction in fibrosis degree， and significant reductions in the positive areas of α-SMA and COL1A1 （P<0.001） and the protein expression levels of α-SMA， TβRⅡ， and p-Smad2/3 （P<0.01）. ConclusionKL7 has a significant targeted therapeutic effect on pancreatic fibrosis in CP mice through specific binding of KL7 to TβRⅡ， thereby inhibiting the activation of the TGF-β/Smad signaling pathway.

Objective To investigate the role of cholinergic receptor muscarinic 3(Chrm3)in regulating lipopolysaccharide(LPS)-induced inflammation in peritoneal macrophages in lipopolysaccharide binding protein(LBP)-knockout(Lbp-/-)mice.Methods Peritoneal macrophages were isolated from wild-type and Lbp-/-mice to establish an LPS-induced inflammation model.Chrm3 expression in Lbp-/-mouse peritoneal macrophages was inhibited by 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide(4-damp)and small interfering(siRNA)and Chrm3 overexpression was achieved by lentivirus transfection.For 4-damp inhibition,cells were divided into control,LPS,and inhibitor groups,and for siRNA transfection,cells were divided into control,LPS,si-normal control group,and si-Chrm3 groups.For overexpression,cells were divided into control,LPS,negative control,and overexpression groups.Changes in Chrm3 in response to LPS stimulation were verified by Western blot.The effects of 4-damp,si-Chrm3,and lentivirus on cell inflammation and survival were confirmed by Cell Counting Kit-8,quantitative polymerase chain reaction,and Western blot assays.Results Chrm3 protein expression was significantly elevated in Lbp-/-peritoneal macrophages post-LPS stimulation(P＜0.001),whereas there was no notable change in wild-type cells.The cell survival rate was significantly increased in the 4-damp and si-Chrm3 groups(P＜0.05,P＜0.01),and cell survival was significantly reduced in the overexpression group(P＜0.01).Furthermore,4-damp and si-Chrm3 significantly reduced expression levels of the inflammatory factors tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6(P＜0.01,P＜0.001),and phospho-extracellular signal-regulated kinase(p-ERK)(P＜0.01,P＜0.001),which are associated with cell damage and inflammation.In contrast,TNF-α,IL-1β,IL-6(P＜0.001),and p-ERK protein(P＜0.001)were significantly elevated in the overexpression group.Conclusions LPS stimulation upregulated the expression of Chrm3 and proinflammatory cytokines in Lbp-/-peritoneal macrophages.Specific downregulation of Chrm3 by 4-damp and si-Chrm3 significantly decreased LPS-induced proinflammatory cytokines in Lbp-/-peritoneal macrophages,while upregulation of Chrm3 using overexpressing lentivirus significantly elevated the expression of related inflammatory factors.Chrm3 is implicated in the regulation of the LPS-induced inflammation response in peritoneal macrophages in Lbp-/-mice.

[Objective]To discuss the clinical manifestations and image features of Neuromyelitis Optica Spectrum Disorder(NMOSD)with respiratory insufficiency.We present a retrospective review about the use of double filtration plasmapheresis in the treatment of the acute attack of NMOSD in these patients.[Methods]All of our patients with central respiratory insufficiency who suffered attacks of NMOSD were retrospectively considered for inclusion.Extended Disability Status Scale(EDSS)scores were compared within six months after double membrane filtration plasma exchange.[Results]The clinical data of the six patients included were analyzed.Magnetic Resonance Imaging confirmed that the demyelinating plaques in our patients could involve the medulla oblongata and upper spinal cord.They were managed by plasma exchange given as an initial therapy.The clinical symptoms improved significantly and the patients were successfully withdrawn from the ventilator,with EDSS scores significantly reduced(P<0.001).[Conclusion]Demyelination of medulla oblongata and upper spinal cord in NMOSD may lead to acute life-threatening respiratory compromise,and early initiation of double filtration plasmapheresis can be a safe and effective treatment.

Objective To investigate the role of cholinergic receptor muscarinic 3(Chrm3)in regulating lipopolysaccharide(LPS)-induced inflammation in peritoneal macrophages in lipopolysaccharide binding protein(LBP)-knockout(Lbp-/-)mice.Methods Peritoneal macrophages were isolated from wild-type and Lbp-/-mice to establish an LPS-induced inflammation model.Chrm3 expression in Lbp-/-mouse peritoneal macrophages was inhibited by 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide(4-damp)and small interfering(siRNA)and Chrm3 overexpression was achieved by lentivirus transfection.For 4-damp inhibition,cells were divided into control,LPS,and inhibitor groups,and for siRNA transfection,cells were divided into control,LPS,si-normal control group,and si-Chrm3 groups.For overexpression,cells were divided into control,LPS,negative control,and overexpression groups.Changes in Chrm3 in response to LPS stimulation were verified by Western blot.The effects of 4-damp,si-Chrm3,and lentivirus on cell inflammation and survival were confirmed by Cell Counting Kit-8,quantitative polymerase chain reaction,and Western blot assays.Results Chrm3 protein expression was significantly elevated in Lbp-/-peritoneal macrophages post-LPS stimulation(P＜0.001),whereas there was no notable change in wild-type cells.The cell survival rate was significantly increased in the 4-damp and si-Chrm3 groups(P＜0.05,P＜0.01),and cell survival was significantly reduced in the overexpression group(P＜0.01).Furthermore,4-damp and si-Chrm3 significantly reduced expression levels of the inflammatory factors tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6(P＜0.01,P＜0.001),and phospho-extracellular signal-regulated kinase(p-ERK)(P＜0.01,P＜0.001),which are associated with cell damage and inflammation.In contrast,TNF-α,IL-1β,IL-6(P＜0.001),and p-ERK protein(P＜0.001)were significantly elevated in the overexpression group.Conclusions LPS stimulation upregulated the expression of Chrm3 and proinflammatory cytokines in Lbp-/-peritoneal macrophages.Specific downregulation of Chrm3 by 4-damp and si-Chrm3 significantly decreased LPS-induced proinflammatory cytokines in Lbp-/-peritoneal macrophages,while upregulation of Chrm3 using overexpressing lentivirus significantly elevated the expression of related inflammatory factors.Chrm3 is implicated in the regulation of the LPS-induced inflammation response in peritoneal macrophages in Lbp-/-mice.

Objective To observe the effect of acupuncture synchronized with speech training on speech function of patients with post-stroke motor aphasia.Methods Sixty inpatients with post-stroke motor aphasia were selected from the Affiliated Hospital of Shandong Univer-sity of Traditional Chinese Medicine,from January to August,2023.They were randomly divided into control group(n=30)and synchronous group(n=30).Both groups received acupuncture and speech training;the con-trol group received acupuncture in the morning and speech training in the afternoon,while the synchronous group received acupuncture and speech training synchronously,for three weeks.They were assessed with Chi-nese Rehabilitation Research Center Standard Aphasia Examination(CRRCAE),Non-Language-Based Cognitive Assessment(NLCA)and Communication Activities of Daily Living(CADL)before and after treatment.Results The subscores of CRRCAE and NLCA,and score of CADL increased in both groups(|t|>2.081,P<0.05)after treatment,and they were better in the synchronous group than in the control group(|t|>2.680,P<0.05).Conclusion Synchronous mode of acupuncture and speech training is more effective on post-stroke motor aphasia than time-sequence mode.

Objective To observe the effect of synchronous acupuncture and articulation training on spastic dysarthria after stroke.Methods From January to August,2023,64 stroke patients in the Affiliated Hospital of Shandong University of Tradi-tional Chinese Medicine were selected,and randomly divided into control group(n=32)and synchronous group(n=32).Both groups received routine neurological treatment and basic articulation training.The control group added asynchronous acupuncture,while the synchronous group added synchronous acupuncture,for four weeks.Before and after treatment,the modified Frenchay Dysarthria Assessment(m-FDA),Speech Intelligibility Test(SIT),maximum phonation time(MPT)and maximum counting ability(MCA)were used to evaluate the curative effects.Results After treatment,the scores of m-FDA in all the dimensions decreased in both groups(t>2.882,P<0.01);ex-cept for the jaw dimension,the scores of m-FDA in reflexes,respiration,lips,soft palate,larynx,tongue and speech dimensions were significantly lower in the synchronous group than in the control group(t>2.050,P<0.05).After treatment,the results of SIT,and MPT and MCA significantly increased in both groups(t>21.061,P<0.001),and they were better in the synchronous group than in the control group(t>11.412,P<0.001).Conclusion Synchronous acupuncture and articulation training can effectively alleviate the severity of spastic dysarthria after stroke,which was superior to asynchronous acupuncture and articulation training.

Objective To explore the characteristic changes in white matter microstructure in Parkinson's disease(PD)patients via automated fiber quantification(AFQ)technology,providing a basis for the identification and diagnosis of PD,and to analyze the feasibility of combining the AFQ method with support vector machine(SVM)in the diagnosis of PD.Methods Forty patients with primary PD(PD group)and 20 healthy controls(HC)(HC group)were prospectively selected.The AFQ technology was applied for white matter fiber tract analysis.Statistical analyses were performed using FSL(v6.0)software and SPSS 27.0 software.Independent-sample t-tests were conducted for comparisons between groups in AFQ analysis.The AFQ method was used to analyze the relationship between diffusion tensor imaging(DTI)parameters and Montreal Cognitive Assessment(MoCA)scores.Results(1)The results of AFQ analysis revealed that compared with the HC group,the PD group exhibited significantly lower fractional anisotropy(FA)values in the right cingulum bundle,left cingulum bundle hippocampus,and left uncinate fasciculus,with no differences in the FA values of the remaining 17 fiber tracts.Moreover,PD group demonstrated higher mean diffusivity(MD)values in the left cingulum bundle,left cingulum bundle hippocampus,left inferior frontal occipital fasciculus,left inferior longitudinal fasciculus,left superior longitudinal fasciculus,and left uncinate fasciculus.These differences were statistically significant(P<0.05),while no significant differences were found in the MD values of the remaining 14 fiber tracts.Furthermore,the MD values of the left inferior frontal occipital fasciculus,and left inferior longitudinal fasciculus were negatively correlated with the MoCA scores.(2)The classification results of SVM showed that the best results were achieved when combining the differential nodes of FA and MD as classification features,with an area under the curve(AUC)of 0.922,an accuracy of 84.81%,a sensitivity of 87.50%,and a specificity of 82.05%.Conclusion The DTI parameters in PD patients can serve as potential biomarkers for diagnosis.The AFQ methods provides an effective approach for detecting alterations white matter tract integrity,offering important insights for the identification and diagnosis of PD.The best results are achieved when combining the differential nodes of FA and MD as classification features.

Objective To investigate the effect of ClearInfinity deep learning reconstruction algorithm on image quality and radiation dose of abdominal computed tomography angiography(CTA)at low kV and low contrast medium.Methods One hundred patients who underwent abdominal CTA were selected and randomly divided into group A and group B.Group A:tube voltage 70 kV,con-trast medium 30-35 mL,divided into A1 and A2 subgroups according to reconstruction algorithm,group A1 50%ClearInfinity,group A2 50%ClearView iterative algorithm;group B:tube voltage 100 kV,contrast medium 60-70 mL,50%ClearView.CT values and standard deviation(SD)values of region of interest(ROI)of abdominal aorta,proper hepatic artery,superior mesenteric artery,renal artery and common iliac artery were evaluated objectively,while signal-to-noise ratio(SNR)and contrast-to-noise ratio(CNR)were calculated;subjective scores were evaluated by two physicians;radiation doses of groups A and B were analyzed.Results Volume CT dose index(CTDIvol),dose length product(DLP)and effective dose(ED)in group A were significantly lower than those in group B(P<0.05),subjective scores in group A1 and group B were higher than those in group A2(P<0.05),and there was no difference between group A1 and group B(P>0.05).Compared with group A1,SNR and CNR of all vessels in group A2 were significantly decreased.CT values of abdominal aorta and common iliac artery,CNR of common iliac artery and supe-rior mesenteric artery in group B were significantly increased,SNR of renal artery was significantly decreased(P<0.05).Conclusion ClearInfinity deep learning reconstruction algorithm combined with 70 kV scanning technology can obtain better abdom-inal CTA image quality,and effectively reduce the radiation dose and contrast medium of patients,which has high clinical application value.

Bronchial asthma(hereinafter referred to as asthma)is a common chronic respiratory disease characterized by airway inflammation,airway hyperresponsiveness,and airway remodeling.Its pathogenesis is highly complex and heterogeneous,involving multiple factors such as genetics,immunity,and environmental exposure.Currently,therapeutic options for asthma remain relatively limited,making it an urgent priority to explore its underlying mechanisms,identify effective treatment strategies,and develop new drugs.In this context,the establishment of animal models for asthma plays an irreplaceable and crucial role.However,to date,no single ideal animal model has been able to fully and accurately replicate all the features of the onset and progression of human asthma.This study systematically reviews the research progress over the past five years in the establishment methods of asthma animal models.It provides a detailed overview of commonly used experimental animals(such as mice,rats,and guinea pigs),frequently used sensitizing agents(including ovalbumin,house dust mite,lipopolysaccharide,and toluene diisocyanate),and the methods for establishing asthma models using these animals and sensitizers.This study also presents an objective evaluation of the advantages,limitations,and applicability of each model.Evaluation criteria for asthma models are summarized across multiple dimensions,including behavioral assessments,pulmonary function,histopathology,immunological indicators,and pharmacodynamics.Although methods for establishing refractory asthma models remain underdeveloped,several strategies for modeling refractory asthma have been summarized through a review of relevant literature,aiming to provide useful references for related research.Based on current scientific and technological advancements,it is anticipated that future research on asthma animal models will focus more on clinical relevance,technological innovation,and multidisciplinary integration.Specifically,future models are expected to adopt multi-sensitizer induction protocols,apply cutting-edge tools such as gene editing,enhance clinical relevance and promote diversification and personalization of models.Furthermore,advanced technologies such as bioimaging and biosensing are anticipated to enable dynamic monitoring of airway inflammation and remodeling.Organ-on-a-chip platforms may also be explored as potential alternatives to traditional animal models.The ultimate goal is to develop multifactorial,composite models that better simulate the complexity and heterogeneity of human asthma.