ObjectiveTo explore the potential mechanism of Xianglian Huazhuo prescription （XLHZ） in treating chronic atrophic gastritis （CAG） by regulating cell cycle and inhibiting proliferation， using bioinformatics technology and animal experiments. MethodsDifferential expressed genes （DEGs） related to CAG were screened using GEO database and GEO2R tool. Weighted gene co-expression network analysis （WGCNA） was employed to search for hub genes of CAG. These hub genes were intersected with cell cycle proliferation based on GeneCards database. Eenrichment analysis of the intersecting genes was performed to obtain signaling pathways and biological processes related to CAG. Protein protein interaction （PPI） analysis of genes was conducted using the Protein Interaction Platform （STRING） database to search the super hub gene （hub 2.0）， and animal experiments were conducted for further validation. Fourteen of 70 male Wistar rats were randomly selected as the normal group， and the remaining 56 rats were prepared by the combined modeling method of "starvation disorder+N-methyl-N-nitro-N-nitrosoguanidine （MNNG） + sodium salicylate". The successfully modeled rats were randomly divided into the model group， XLHZ-H， XLHZ-M， and XLHZ-L groups （36， 18， 9 g·kg^-1， respectively）， and Morodan group （1.4 g·kg^-1）. Each group was given corresponding intervention for 60 days. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes of gastric mucosa in rats. The ultrastructure of gastric mucosal tissue cells was observed by transmission electron microscopy. The relative expression levels of TGF-β₁， Smad2 and Smad3 proteins， S/G₂/M phase marker geminin and proliferation marker MCM2 were detected by Western blot in gastric mucosal tissue， and Spearman correlation analysis was performed. ResultsA total of 15 hub 2.0 genes were identified， including TGF-β₁， suggesting the involvement of the TGF-β₁ signaling pathway in the CAG pathogenesis. Compared with the normal group， the expressions of TGF-β₁， Smad2， geminin and MCM2 proteins in the gastric mucosa tissue of the model group were increased （P<0.05）， and the expression of Smad3 protein was decreased （P<0.05）. Compared with the model group， the expressions of TGF-β₁ and geminin in the gastric mucosa were decreased in the drug groups （P<0.05）. The XLHZ-M group， XLHZ-H group and Morodan group had significantly decreased protein expression of Smad2 and MCM2 （P<0.05）. The protein expression of Smad3 was significantly increased in XLHZ-M， XLHZ-H， and Morodan groups （P<0.05）. Spearman correlation analysis showed that Smad3 was negatively correlated with other indicators， and positively correlated with other indicators （P<0.01）. ConclusionXLHZ may inhibit TGF-β₁/Smads signaling pathway， regulate cell cycle， and inhibit proliferation in the treatment of CAG.

ObjectiveTo explore the potential mechanism of Xianglian Huazhuo prescription （XLHZ） in treating chronic atrophic gastritis （CAG） by regulating cell cycle and inhibiting proliferation， using bioinformatics technology and animal experiments. MethodsDifferential expressed genes （DEGs） related to CAG were screened using GEO database and GEO2R tool. Weighted gene co-expression network analysis （WGCNA） was employed to search for hub genes of CAG. These hub genes were intersected with cell cycle proliferation based on GeneCards database. Eenrichment analysis of the intersecting genes was performed to obtain signaling pathways and biological processes related to CAG. Protein protein interaction （PPI） analysis of genes was conducted using the Protein Interaction Platform （STRING） database to search the super hub gene （hub 2.0）， and animal experiments were conducted for further validation. Fourteen of 70 male Wistar rats were randomly selected as the normal group， and the remaining 56 rats were prepared by the combined modeling method of "starvation disorder+N-methyl-N-nitro-N-nitrosoguanidine （MNNG） + sodium salicylate". The successfully modeled rats were randomly divided into the model group， XLHZ-H， XLHZ-M， and XLHZ-L groups （36， 18， 9 g·kg^-1， respectively）， and Morodan group （1.4 g·kg^-1）. Each group was given corresponding intervention for 60 days. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes of gastric mucosa in rats. The ultrastructure of gastric mucosal tissue cells was observed by transmission electron microscopy. The relative expression levels of TGF-β₁， Smad2 and Smad3 proteins， S/G₂/M phase marker geminin and proliferation marker MCM2 were detected by Western blot in gastric mucosal tissue， and Spearman correlation analysis was performed. ResultsA total of 15 hub 2.0 genes were identified， including TGF-β₁， suggesting the involvement of the TGF-β₁ signaling pathway in the CAG pathogenesis. Compared with the normal group， the expressions of TGF-β₁， Smad2， geminin and MCM2 proteins in the gastric mucosa tissue of the model group were increased （P<0.05）， and the expression of Smad3 protein was decreased （P<0.05）. Compared with the model group， the expressions of TGF-β₁ and geminin in the gastric mucosa were decreased in the drug groups （P<0.05）. The XLHZ-M group， XLHZ-H group and Morodan group had significantly decreased protein expression of Smad2 and MCM2 （P<0.05）. The protein expression of Smad3 was significantly increased in XLHZ-M， XLHZ-H， and Morodan groups （P<0.05）. Spearman correlation analysis showed that Smad3 was negatively correlated with other indicators， and positively correlated with other indicators （P<0.01）. ConclusionXLHZ may inhibit TGF-β₁/Smads signaling pathway， regulate cell cycle， and inhibit proliferation in the treatment of CAG.

Purpose To evaluate the static structural changes of mitral annulus and leaflets in mitral valve prolapse via three-dimensional transesophageal echocardiography(3D-TEE),and their continuous dynamic changes during the cardiac cycle.Materials and Methods Twenty-six patients with mitral valve prolapse diagnosed by transesophageal echocardiography in Wuhan Asia Heart Hospital from June 2022 to June 2023 were retrospectively included,and 30 patients who were examined by transesophageal echocardiography in our hospital during the same period due to monitoring of patent foramen ovale or non-cardiac surgery were selected as the control group.The related parameters of the valve annulus were measured.In addition,the meaningful parameters were measured at late diastole(LD),early systole(ES),middle systole(MS)and late systole(LS),and the change rate was calculated.The static structure of mitral valve device was compared between the two groups,and the differences of various parameters at different times were analyzed.Results Compared with the control group,the annulus height(AH)and the ratio of annulus height and combined width(AHCWR)in the case group decreased,and the other parameters increased(t=-7.447-12.472,all P<0.05),but there was no statistical difference in the maximum displacement distance of the annulus between the two groups(P>0.05).Comparing the related parameters of LD,ES,MS and LS at four different time points,compared with the control group,all the parameters in the case group increased except AH and AHCWR(t=-16.224-15.683,all P<0.05).Comparing the continuous changes at four different times,the continuous changes of all parameters in the control group were statistically significant(F=7.632-323.837,all P<0.05),and there were significant differences in the parameters of the control group from LD to ES(all P<0.05).Except anterolateral to posteromedial diameter,intertrigonal diameter and annulus area,the continuous changes of other parameters in the case group were statistically significant(F=3.041-71.082,all P<0.05),and only non-planar angle,AH and AHCWR in the case group had statistical differences in LD to ES(all P<0.05).Compared with the control group,the change rate of parameters in the case group decreased,and the difference was statistically significant(t/U=-7.069-780.000,all P<0.05),and the change rate of C-ring was greater than that of intertrigonal diameter.Conclusion 3D-TEE can quantitatively analyze the dynamic changes of mitral annulus and leaflets,and analyze the continuous changes of mitral valve in cardiac cycle in detail,which can provide more accurate preoperative evaluation for patients and guide treatment.

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 relationship between ribonuclease A3(RNase3)gene polymorphism and its expression level with airway inflammation and glucocorticoid efficacy in children with bronchial asthma(BA).Methods A total of 110 children with bronchial asthma admitted to Handan Maternal and Child Health Hospital June 2022 to June 2024 were selected as the study objects.According to the severity of the condition at admission,they were divided into mild group(n=64),moderate to severe group(n=46),and children who underwent physical examination in the hospital during the same period were selected as the control group(n=82).Molecular weight array gene analysis(MassArray)was used to detect the genotyping of RNase3 gene rs2073342 locus.Quantitative real-time PCR(RT-PCR)was used to detect the expression level of RNase3 mRNA.The clinical data of the children were collected,and the differences of genotype and allele frequency were compared.The relationship between RNase3 gene polymorphism and BA susceptibility was analyzed by unconditioned Logistic regression.Results Compared with the mild group,IL-6(21.49±3.01ng/L vs 13.21±2.84ng/L)and PCT(19.16±4.02μg/L vs 9.94±3.15μg/L)in the moderate and severe group were significantly increased(t=-14.568,-12.952),while FVC,PEF and FEV1 were significantly decreased(t=2.534,3.304,2.011),and the differences were statistically significant(all P<0.05).The genotype distribution of RNase3 gene rs2073342 in both control and study groups was consistent with Hardy-Weinberg balance law(χ2=0.402,0.689,all P>0.05),indicating population representation.Compared with the control group and the mild group,the CC,CA genotype frequencies were higher in moderate to severe group(χ2=35.008,23.079),Compared with the control group,the frequency of CC and CA genotypes in mild group was higher(χ2=7.325),the differences were statistically significant(all P<0.05).Compared with mild group,RNase3 mRNA expression in peripheral blood mononuclear cells of children with C gene BA at rs2073342 in moderate to severe group was higher,and the difference was statistically significant(t=-19.622,P<0.05).In the same group,the mRNA expression level of RNase3 in peripheral blood mononuclear cells of the children with C gene BA at rs2073342 was higher than that of the children without C gene(t=4.169,22.608,all P<0.05).Unconditional Logistic regression results showed that RNase3 gene carrying allele C or dominant model(CC vs CA+AA)was a risk factor for severe disease in BA children(P<0.05).The total effective rate of genotype AA carriers was the highest after glucocorticoid therapy,and the therapeutic effect of genotype AA carriers was significantly better than that of genotype AC and CC,and the difference was statistically significant(χ2=11.858,P<0.05).Conclusion The expression of RNase3 mRNA in peripheral blood of BA children increased significantly with the exacerbation of the disease.Carrier of allele C or dominant model(CC vs CA+AA)is a risk factor for severe disease in BA children.Genotype AA carriers had better therapeutic effect.

Objective To investigate the expression of regional cerebral oxygen saturation(rSO2)in preterm infants with brain injury(BIPI)and its relationship with amplitude-integrated electroencephalogram(aEEG)parameters and neurodevelopment.Methods Retrospectively,116 cases of BIPI(Bilateral Intra-ventricular Perforation)born in the hospital between February 2022 and February 2023 were selected as the study subjects.According to the Expert Consensus on the Diagnosis and Prevention of Premature Infant Brain Injury,these cases were categorized into mild,moderate,and severe groups,with 59,35,and 22 cases,respectively.The study compared the general characteristics of premature infants with different brain injury levels.Regression analysis was conducted to identify the factors affecting brain injury in premature infants,and correlation analyses were performed on aEEG scores,rSO2,and GMS scores.ROC curve analysis was used to evaluate the value of combining brain oxygen saturation monitoring with aEEG in predicting abnormal whole-body motor quality assessment.Results On the 3rd and 7th days after birth,severe brain-injured premature infants had significantly lower rSO2 and aEEG scores compared to those with mild and moderate brain injuries(P<0.05).At 7 days,these infants also had lower scores for graph continuity,sleep-wake cycles,lower boundary amplitude,bandwidth,total score,rSO2,gestational age,and birth weight,and a higher proportion of abnormal developmental outcomes(PR+CS)(P<0.05).Multivariate logistic regression analysis revealed that the 3-day aEEG total score(OR=0.448,95％CI:0.094～0.890),7-day aEEG total score(OR=0.384,95％CI:0.058～0.726),3-day rSO2(OR=0.574,95％CI:0.398～0.750),and 7-day rSO2(OR=0.431,95％CI:0.115～0.777)were all protective factors for brain injury in premature infants,P<0.05.Correlation analysis showed that brain oxygen saturation was negatively correlated with aEEG scores and overall motor quality.The prediction value for abnormal overall motor quality using 7-day rSO2,aEEG,and the combination of 7-day rSO2 and 7-day aEEG scores was as follows:the positive predictive value for rSO2 was 86.46％,the negative predictive value was 96.85％,the sensitivity was 92.31％,the specificity was 92.21％,and the cut-off value was 0.845;The positive predictive value(PPV)of aEEG was 78.26％,the negative predictive value(NPV)was 96.86％,the sensitivity was 89.74％,and the specificity was 92.21％,with a cut-off value of 0.820.The positive predictive value(PPV)of 7-day rSO2 combined with 7-day aEEG score was 89.04％,the NPV was 96.88％,the sensitivity was 94.90％,and the specificity was 93.51％,with a cut-off value of 0.884,P<0.05.ROC curve analysis showed that the AUC of 7-day rSO2+7-day aEEG score was 0.895,De-long test P<0.005,indicating high predictive value for overall exercise quality.Conclusion Combined with rSO2 and EEG,the prediction of overall motor quality is better,which may have clinical value for excluding high-risk children.

Objective To analyze the value of actin-related protein 2(ACTR2)and DEAD-box RNA helicase 3X-linked(DDX3X)expression in evaluating the severity of Rotavirus gastroenteritis(RVGE)in children.Methods A total of 153 children with rotavirus gastroenteritis admitted to Handan Maternal and Child Health Hospital from September 2022 to September 2024 were selected as the research objects.According to the severity of RVGE,the children with RVGE were divided into mild group(n=60),moderate group(n=71)and severe group(n=22).In addition,60 healthy children were randomly selected as the healthy group.The clinical data of the light group,the medium group and the heavy group were compared.The expression levels of ACTR2 and DDX3X in serum of each group were detected by real-time fluorescence quantitative PCR.ROC curve was used to analyze the evaluation value of combined detection of serum ACTR2 and DDX3X in children with severe RVGE.Results The age of children with RVGE in the severe group was significantly lower than that in the mild group and the moderate group,and the differences were statistically significant(Z=8.307,5.885,all P＜0.001).There were statistically significant differences in dehydration and diarrhea among the three groups(F=9.434,126.080,all P＜0.05).The expression levels of ACTR2(1.20±0.28)and DDX3X(1.22±0.37)in RVGE group were significantly higher than those in control group(1.01±0.02,1.04±0.09),and the differences were statistically significant(t=5.071,3.584,all P＜0.05).The expression levels of ACTR2[1.11±0.23,1.23±0.21,1.42(1.25,1.57)]and DDX3X[1.14±0.22,1.25±0.24,1.32(1.23,1.62)]in the serum of the mild group,the moderate group and the severe group increased in turn,and the differences were statistically significant(Z=27.196,18.013,all P＜0.05).The ROC curve analysis results showed that the combined detection of serum ACTR2 and DDX3X predicted the AUC(95%CI)of critically ill RVGE patients,with higher sensitivity and specificity than the detection of the two alone(Z=2.573,2.101,P=0.014,0.034).Conclusion The expression levels of serum ACTR2 and DDX3X are closely related to the severity of RVGE.The combined detection of the two has a high predictive value for the diagnosis of the severity of RVGE.

BACKGROUND:Studies have shown that immune cells are involved in all processes of ischemic stroke,in which cuproptosis also plays a key role.OBJECTIVE:To screen diagnostic biomarkers related to the progression of ischemic stroke through bioinformatics,and analyze and validate cuproptosis-related genes closely related to the occurrence and development of ischemic stroke.METHODS:The GSE16561 microarray was obtained from the GEO database,containing data from 39 cases of ischemic stroke(ischemic stroke group)and 24 controls(control group).Differentially expressed genes from the ischemic stroke microarray data were analyzed.Gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed.By using LASSO and Random Forest methods,key genes affecting the occurrence and development of ischemic stroke were screened,and a diagnostic model was established and validated.Differential gene analysis was performed through immune cell infiltration and weighted gene co expression network.The differentially expressed immune-related genes were intersected with cuproptosis genes to obtain the hub genes related to cuproptosis immunity.In vitro cell experiments were conducted to divide rat hippocampal neurons into a normal group and an ischemic stroke group,and qPCR experiments were performed to verify the results.RESULTS AND CONCLUSION:(1)573 differentially expressed genes were obtained by differential analysis.Differentially expressed genes were mainly enriched in biological processes,such as positive regulation of immune response,and signaling pathways such as lipid and atherosclerosis.(2)Machine learning methods were used to screen diagnostic genes such as MFN2,PKM2,CREG1,and FOXO3A,which may have some diagnostic value for ischemic stroke.(3)Immune infiltration analysis revealed resting plasma cells,NK cells,macrophages,etc.,indicating that immune cells play a certain role in the pathogenesis of ischemic stroke.(4)Weighted gene co-expression network analysis combined with immune infiltration analysis obtained 118 key module genes,which were intersected with cuproptosis genes to obtain 2 cuproptosis and immune characteristic genes.The correlation analysis between four diagnostic genes and Hub genes showed that the expression of FOXO3A and MFN2,PKM2 and BCL2L1,MTF1 and MFN2,ATP7B and BCL2L1 were correlated.(5)The qPCR results showed significant differences in the genes MTF1 and ATP7B between the ischemic stroke group and the blank group.To conclude,ATP7B and MTF1 can serve as characteristic genes for cuproptosis in ischemic stroke.It is possible to improve ischemic stroke by intervening in ATP7B and MTF1 to regulate cuproptosis.

Objective:To explore the relationship between early fecal calprotectin (FC) level and the long-term efficacy of infliximab (IFX) in the treatment of Crohn′s disease (CD) and predictive the value.Methods:From January 2018 to December 2023, at the First Affiliated Hospital with Nanjing Medical University, the clinical data of patients with moderate-to-severe CD who received IFX as first-line therapy were retrospectively collected. The main outcomes were clinical and endoscopic remission at week 52 after IFX treatment, and the secondary outcome was clinical response at week 14 after IFX treatment. The predictive value of FC levels at week 0 (at baseline when first administered) and week 14 of treatment was evaluated for the clinical and endoscopic remission at week 52 after IFX treatment. Multivariate logistic regression was performed to investigate the factors predicting endoscopic remission. The optimal cutoff value was calculated, model was established, the data was divided into training set and validation set at a ratio of 7∶3 using the random number table method and the corresponding column chart was drawn. Receiver operating characteristic curve (ROC) and calibration curve were used to evaluate the discrimination and calibration of the model, respectively. Mann-Whitney U test was used for statistical comparison. Results:A total of 165 patients with CD were enrolled, of whom 150 cases (90.9%) achieved clinical response after induction therapy, and 15 cases (9.1%) were primary non-response. Among the 150 patients with clinical response, 112 cases (74.7%) achieved clinical remission at week 52 after treatment, while 38 cases (25.3%) did not achieve clinical remission. Endoscopic evaluation was performed at week 52 after treatment in 139 patients, of whom 54 cases (38.8%) achieved endoscopic remission and 85 cases (61.2%) did not. At week 14 of treatment, there was no statistically significant difference in FC level between the patients achieved and did not achieve clinical response (263.24 (93.96, 675.28) μg/g vs. 556.35 (245.77, 953.56) μg/g, P>0.05). At week 52 after treatment, the FC level of patients who achieved clinical remission was lower than that of patients did not achieve(103.20(44.11, 456.57) μg/g vs. 531.26(222.06, 998.40) μg/g) and the decreased value of FC at week 52 and week 0 after treatment of patients achieved clinical remission was more than that of patients did not achieve clinical remission (443.34 (82.25, 788.95) μg/g vs. 269.91 (-79.20, 522.54) μg/g), and the differences were statistically significant ( U=1 078.00, 2 677.00; P<0.001, =0.018). At week 52 after treatment, the FC level of patients achieved endoscopic remission was lower than that of patients did not achieve endoscopic remission (52.80(31.93, 83.47) μg/g vs. 506.18(217.44, 778.02) μg/g), and the decreased value of FC at week 52 and week 0 after treatment of patients achieved endoscopic remission was more than that of patients did not achieve endoscopic remission (428.85(140.20, 863.60) μg/g vs. 309.61(-62.37, 683.82) μg/g), and the differences were statistically significant ( U=500.00, 2 812.00; P<0.001, =0.025). The FC level at week 14 of treatment could predict the clinical and endoscopic remission at week 52 after treatment (area under the curve (AUC) =0.663, 0.773; 95% confidence interval (95% CI): 0.566 to 0.760, 0.694 to 0.852; P=0.006, <0.001). The optimal cutoff value of FC at week 14 of treatment for predicting endoscopic remission at week 52 after treatment was 246.13 μg/g, with a sensitivity of 0.741 and a specificity of 0.671. The results of multivariate logistic regression analysis revealed that FC ≤ 246.13 μg/g at week 14 of treatment ( OR=4.576, 95% CI: 2.021 to 10.363, P<0.001), baseline albumin ( OR=1.093, 95% CI: 1.006 to 1.188, P=0.035), and baseline platelet-to-lymphocyte ratio (PLR) ( OR=0.995, 95% CI: 0.990 to 1.000, P=0.046) were independent influencing factors of endoscopic remission at week 52 after treatment. A predictive model for endoscopic remission at week 52 after IFX treatment was established based on FC ≤ 246.13 μg/g at week 14 of treatment, baseline albumin and PLR. The results of ROC analysis showed that this model had good discriminative ability, with an AUC of 0.780 (95% CI: 0.700 to 0.878) in the validation set, with a sensitivity of 0.812 and a specificity of 0.760. The results of calibration curve analysis demonstrated that the average absolute error of the prediction model in the validation set was 0.038, and the consistency between the predicted probability and the actual probability was good. Conclusion:FC ≤ 246.13 g/g at week 14 of IFX treatment has good predictive value for endoscopic remission at week 52 after treatment in CD patients.