Despite of the widespread use of hydrogen and methane breath test, the variability in testing protocols, gas measurement techniques, and interpretation criteria continues to challenge the reproducibility and comparability across centers, especially in the Asia-Pacific region. The Asian Neurogastroenterology and Motility Association hence presents the first Asian monograph guiding application and interpretation of breath test. The monograph was formulated according to the framework of indications, preparatory process, performance, and interpretation of results, as well as future direction for research.

Background/Aims: Belching can occur either independently or concurrently with reflux symptoms. This study aims to explore the relationship between belching and reflux and explore the underlying mechanisms involved. Methods: Firstly, patients with isolated belching symptoms or concurrent belching and reflux symptoms were retrospectively enrolled. High-resolution esophageal impedance manometry (HRIM) and 24-hour pH impedance monitoring were used to assess belching types and temporal relationship between belching and reflux. Secondly, subjects with belching disorder were prospectively recruited. Postprandial HRIM monitoring was used to clarify the relationship and underlying mechanisms between belching and reflux. Results: In the initial phase, 139 subjects were enrolled. Regardless of patients' symptom spectrum (with or without gastroesophageal reflux symptoms), the proportion of preceding-reflux belching was significantly higher in those with objective evidence of gastroesophageal reflux disease, compared to those without (both P < 0.05), which indicated that belching could trigger reflux events and led to more severe reflux burden. To clarify the potential mechanisms involved, 44 belching subjects were prospectively enrolled in the second phase of this study. HRIM analysis demonstrated that maximum intragastric pressure, gastric–lower esophageal sphincter pressure gradient and gastric–esophageal pressure gradient were significantly higher in preceding-reflux belching than those in independent belching. Conclusions This study demonstrated that belching might trigger reflux by increasing intragastric pressure, elevating the gastric-sphincter pressure gradient, which enables gastric contents to pass through the esophagogastric junction.These findings have crucial clinical significance, suggesting that reducing the occurrence of belching might be a potential treatment for refractory reflux.

ObjectiveTo evaluate the clinical efficacy of Huangqin Qingre Chubi capsules （HQC） in patients with rheumatoid arthritis （RA） and the syndrome of dampness-heat obstruction and investigate the mechanism by which HQC restore clinical and laboratory parameters in RA patients through regulation of the long non-coding RNA （LncRNA） endogenous Bornavirus-like nucleoprotein 3 pseudogene （EBLN3P）/miR-369-3p/nuclear factor I X（NFIX）-Janus kinase（JAK）/signal transducer and activator of transcription （STAT） signaling pathway，thereby providing new experimental evidence and potential therapeutic targets for the traditional Chinese medicine （TCM） treatment of RA. MethodsA randomized controlled trial design was adopted，with 100 RA patients meeting the criteria assigned into an observation group （n=50） and a control group （n=50）. In addition，50 healthy controls were enrolled. The control group received leflunomide plus meloxicam，while the observation group additionally received HQC. Efficacy was assessed after 4 weeks of treatment. Observation indicators including Western medicine indicators （DAS28，CPRI-RA，SF-36，VAS，PGA，PhGA，and SAS/SDS），TCM syndrome scores （TCMSS-RADHP，TCMSS-RASDP），and laboratory indicators （ESR，CRP，RF，anti-CCP antibody，VEGF，bFGF，MMP-2/9，p-JAK2/STAT3，and LncRNA EBLN3P/miR-369-3p/NFIX expression） were measured to comprehensively explore the therapeutic mechanism of HQC. ResultsThe study included 45 healthy controls （34 males and 11 females） and 87 RA patients （22 males and 65 females）. The median age，median disease duration，and median BMI of patients were 56 years old，10 years，and 21.51 kg·m^-2，respectively. Compared with healthy controls，the RA group exhibited a typical state of immune-inflammatory activation as manifested by elevated levels of ESR，anti-CCP antibodies，CRP，and RF （P<0.01），raised clinical symptom scores （DAS28，VAS，PGA，PhGA，CPRI-RA，and TCM symptoms） （P<0.05，P<0.01），and reduced SF-36 domain scores （P<0.01）. Molecular detection revealed abnormal expression of LncRNA EBLN3P and miR-369-3p，upregulated expression of key proteins in the JAK/STAT pathway （JAK2/p-JAK2 and STAT3/p-STAT3） and angiogenesis markers （VEGF，bFGF，and MMP-2/9），and downregulated expression of NFIX （P<0.05，P<0.01） in the RA group. Spearman correlation analysis further demonstrated associations of key molecules in the LncRNA EBLN3P/miR-369-3p/NFIX-JAK/STAT pathway with immune-inflammatory markers and clinical symptom scores （P<0.05，P<0.01）. Regarding treatment response，both groups showed reductions in ESR，anti-CCP，CRP，and RF levels compared with baseline （P<0.01） and restored scores of DAS28，CPRI-RA，VAS，PGA，PhGA，TCM symptoms，SAS，SDS，and all SF-36 domains （P<0.01）. The observation group outperformed the control group in restoring the scores of SAS，SDS，PGA，TCMSS-SDP，and SF-36 domains （PF，BP，GH，VT，and SF） （P<0.05）. Real-time PCR analysis revealed that post-treatment expression levels of LncRNA EBLN3P，miR-369-3p，NFIX，JAK2，p-JAK2，STAT3，p-STAT3，VEGF，bFGF，MMP-2，and MMP-9 were recovered compared with pre-treatment levels （P<0.01）. Notably，the observation group demonstrated superior regulatory effects on p-JAK2，STAT3，p-STAT3，and MMP-9 compared with the control group （P<0.05）. Apriori association rule analysis revealed that HQC treatment was significantly correlated with reduced TCMSS-RADHP，ESR，SF，DAS28，SAS，and SDS scores，as well as increased VT，GH，and RP scores （confidence >80%，support >50%）. Regarding safety，two patients in the observation group experienced mild gastrointestinal discomfort，and one patient in the control group developed a rash. All the symptoms resolved after symptomatic treatment. ConclusionHQC target and regulate the LncRNA EBLN3P/miR-369-3p/NFIX-JAK/STAT axis to inhibit angiogenesis and inflammatory responses，reduce immune inflammatory markers，alleviate clinical symptoms，and improve the quality of life in RA patients，exerting synergistic effects on symptoms and quality of life.

Objective:To discuss the effects of bone marrow-derived mesenchymal stem cells(BMSCs)transplantation on mitophagy in the vascular dementia(VaD)rats through reactive oxygen species(ROS)/nuclear factor erythroid 2-related factor 2(Nrf2)signaling,and to clarify its mechanism.Methods:Forty-five male adult SD rats were randomly divided into sham operation group,model group,unloaded group,BMSCs group,and MSCs+ML385(Nrf2 inhibitor)group(combination group),and there were 9 rats in each group.After intraperitoneal anesthesia,the VaD models were established in all groups except sham operation group.Morris water maze test was used to detect the learning and memory abilities of the rats in various groups;HE staining was used to observe the histopathological morphology of brain tissue of the rats in various groups;Nissl staining was used to observe the changes of Nissl bodies in hippocampus region of brain tissue of the rats in various groups;transmission electron microscope was used to observe the ultrastructure of hippocampus region of the rats in various groups;fluorescence probe method was used to detect the ROS levels in hippocampus neurons in various groups;Western blotting method was used to detect the expression levels of Nrf2,heme oxygenase-1(HO-1),PTEN-induced putative kinase 1(PINK1),parkin RBR E3 ubiquitin protein ligase(Parkin),Beclin-1,ubiquitin-binding protein p62(P62),and microtubule-associated protein 1A/1B-light chain 3(LC3-Ⅱ/LC3-Ⅰ)ratio in brain tissue of the rats in various groups.Results:The Morris water maze results showed that compared with sham operation group,the escape latency of the rats in model group was significantly increased(P<0.01),while the number of crossing time and residence time were significantly decreased(P<0.01).Compared with model group,the escape latency of the rats in BMSCs group was significantly decreased(P<0.01),while the number of crossing time and residence time were significantly increased(P<0.01).Compared with BMSCs group,the escape latency of the rats in combination group was significantly increased(P<0.01),while the number of crossing time and residence time were significantly decreased(P<0.01).The HE staining results showed that hippocampus neurons of the rats in sham operation group were normal in quantity and morphology,with uniform staining and clear structure.Compared with sham operation group,the hippocampus tissue of the rats in model group showed sparse arrangement,disordered structure,reduced neuronal quantity,varied morphology,uneven staining,nuclear pyknosis,and partial neuronal necrosis.Compared with model group,the neuronal damage of the rats in hippocampus regio in BMSCs group was alleviated,with restored morphology and improved neuronal loss.Compared with BMSCs group,the neurons of the rats in hippocampus region in combination group showed irregular morphology,disordered structure,unclear cell boundaries,uneven staining,and nuclear pyknosis.The Nissl staining results showed that the hippocampal neurons in sham operation group were tightly arranged with intact morphology,obvious nucleoli,and abundant darkly stained Nissl bodies.Compared with sham operation group,the neurons in hippocampus region of the rats in model group showed pyknosis,vacuolization,and sparse Nissl bodies.Compared with model group,the BMSCs group showed reduced neuronal pyknosis,relatively intact morphology,and increased Nissl bodies.Compared with BMSCs group,the combination group showed neuronal pyknosis,loss of morphological integrity,and fragmented Nissl bodies.The transmission electron microscope results showed that mitochondria in sham operation group exhibited oval shape with intact double-membrane structure and cristae.Compared with sham operation group,the mitochondria in model group showed swelling,disrupted membranes,broken cristae,and numerous autophagosomes.Compared with model group,the BMSCs group showed improved mitochondrial structure and reduced autophagosomes.Compared with BMSCs group,the combination group showed swollen mitochondria,disrupted membranes,broken cristae,and visible autophagosomes.The fluorescence probe results showed that compared with sham operation group,the ROS levels in the hippocampus neurons in brain tissue of the rats in model group were significantly increased(P<0.01);compared with model group,the ROS levels in hippocampus neurons in brain tissue of the rats in BMSCs group were significantly decreased(P<0.01);compared with BMSCs group,the ROS levels in hippocampus neurons in brain tissue of the rats in combination group were significantly increased(P<0.01).The Western blotting results showed that compared with sham operation group,the expression levels of Nrf2 and HO-1 proteins in brain tissue of the rats in model group were significantly decreased(P<0.01);compared with model group,the expression levels of Nrf2 and HO-1 proteins in brain tissue of the rats in BMSCs group were significantly increased(P<0.01);compared with BMSCs group,the expression levels of Nrf2 and HO-1 proteins in brain tissue of the rats in combination group were significantly decreased(P<0.01);compared with sham operation group,the expression levels of Parkin,PINK1,and Beclin-1 proteins,and LC3-Ⅱ/LC3-Ⅰ ratio of the rats in model group were significantly increased(P<0.01),while the expression level of P62 protein was significantly decreased(P<0.01);compared with model group,the expression levels of Parkin,PINK1,and Beclin-1 proteins,as well as the LC3-Ⅱ/LC3-Ⅰ ratio,of the rats in BMSCs group were significantly decreased(P<0.01),while the expression level of P62 protein was significantly increased(P<0.01);compared with BMSCs group,the expression levels of Parkin,PINK1,and Beclin-1 proteins,as well as the LC3-Ⅱ/LC3-Ⅰ ratio,of the rats in combination group were significantly increased(P<0.01),while the expression level of P62 protein was significantly decreased(P<0.01).Conclusion:BMSCs can alleviate the hippocampal neuronal pathological changes and improve cognitive function in the VaD rats,and its mechanism may be related to the regulation of ROS/Nrf2 signaling pathway to inhibit mitophagy.

Objective To observe the effects of Ditan Yizhi Decoction on Wnt/β-catenin signaling pathway and mitochondrial autophagy in vascular dementia(VD)rats.Methods Totally 70 male SD rats were randomly selected as the sham-operation group,with only the common carotid artery isolated and not ligated,and the rest of the rats were used to prepare a rat model of VD by the modified bilateral common carotid artery permanent ligature method.The successfully modeled rats were randomly divided into model group,donepezil group,agonist group and Ditan Yizhi Decoction low-,medium-and high-dosage groups and were given corresponding interventions for 4 weeks.Morris water maze experiment was used to assess the learning and memory ability of rats,HE staining and Nissl staining were used to observe the morphology of hippocampal tissue,transmission electron microscopy was used to observe the ultrastructure of hippocampal neuron,Western blot was used to detect the expressions of PINK1,Parkin,p62,Beclin-1,LC3BⅡ/Ⅰ,β-catenin,GSK-3β,p-GSK-3β protein.Results Compared with the sham-operation group,rats in the model group had a prolonged escape latency(P<0.01),a reduced number of crossing platforms(P<0.01);neuronal gaps in the CA1 area of the hippocampus were enlarged,with irregular cell morphology and blurred borders,neuronal solidification,Nissl bodies dissolution and fragmentation,reduced quantity,mitochondrial cristae disorder,and disruption of the structure of the bilayer membrane,with a greater number of autophagosomes in the surroundings;the expressions of PINK1,Parkin,Beclin-1,LC3BⅡ/I,β-catenin,p-GSK-3β/GSK-3β protein were significantly increased(P<0.01),and p62 protein expression was significantly decreased(P<0.01).Compared with the model group,the escape latency of rats in donepezil group,agonist group and Ditan Yizhi Decoction low-,medium-and high-dosage groups were significantly shortened(P<0.01),and the number of crossing platforms significantly increased(P<0.05,P<0.01);the number of neurons in the hippocampal CA1 region increased,with normal morphology and orderly arrangement,Nissl bodies were abundant,and transmission electron microscopy showed that mitochondrial morphology recovered,rupture decreased,and autophagosomes around mitochondria decreased,the protein expressions of PINK1,Parkin,Beclin-1,LC3BⅡ/Ⅰ,β-catenin and p-GSK-3β/GSK-3β in hippocampal tissue were significantly decreased(P<0.01),the protein expression of p62 significantly increased(P<0.05,P<0.01).Conclusion Ditan Yizhi Decoction can improve learning memory ability and neuronal morphology in VD rats,and its mechanism may be related to the activation of Wnt/β-catenin signaling pathway to inhibit mitophagy.

Objective:This study aimed to investigate the characteristic expression of programmed death-ligand 2(PD-L2)and epithelial-mesenchymal transition(EMT)-related biomarkers in head and neck squamous cell carcinoma(HNSCC)and their mechanism of action in HNSCC metastasis.Methods:Tumor tissue samples from 94 patients with HNSCC were collected from Tianjin Medical University Cancer Hos-pital from January 2018 to July 2023,and their correlation with clinicopathological parameters,including lymph node metastasis,was statist-ically assessed.Western blot was used to detect PD-L2 expression in HNSCC tumor tissues.PD-L2 overexpression and knockdown stably-transfected monoclonal cell lines were generated using lentiviral vectors.Transwell assays were performed to explore the effect of PD-L2 on the invasive migration of the HNSCC cell lines.Additionally,RNA sequencing was used to identify downstream target genes regulated by PD-L2.The expression levels of key EMT markers,including E-cadherin,N-cadherin,and Vimentin,were examined by Western blot to elucidate the molecular mechanism by which PD-L2 promotes tumor cell metastasis through EMT pathway activation.Additionally,RNA sequencing was employed to identify downstream target genes regulated by PD-L2.The expression levels of key EMT markers,including E-cadherin,N-cadherin,and Vimentin,were examined with Western blot analysis to elucidate the molecular mechanism by which PD-L2 promotes tumor cell metastasis through the EMT pathway activation.Results:High expression of PD-L2 is positively correlated with N staging(P<0.05),and elevated PD-L2 expression predicts a poor prognosis in HNSCC patients.Conclusions:PD-L2 regulates the EMT pathway to promote HNSCC metastasis.Targeting PD-L2 is expected to provide a new strategy for the treatment of metastatic HNSCC.

Objective To observe the effects of Ditan Yizhi Decoction on mitochondrial dynamics-mediated glucose metabolism in vascular dementia(VaD)rats based on the ROS/Drp1 axis;To explore its mechanism in treating VaD.Methods Ten male SD rats were randomly selected from 70 as the sham-operation group,and VaD models were prepared using the modified bilateral common carotid artery permanent ligation method for the remaining rats.The successfully modeled rats were randomly divided into model group,positive drug group(donepezil hydrochloride),inhibitor group(Mdivi-1)and Ditan Yizhi Decoction low-,medium-and high-dosage groups(12.86,25.725,51.45 g/kg),and intervened with corresponding method for 4 consecutive weeks.Morris water maze experiment was used to assess the learning memory ability of rats,HE and Nissl staining were used to observe the morphology of hippocampal tissue,transmission electron microscopy was used to observe the mitochondrial ultrastructure of hippocampal neurons,DHE fluorescent probe was used to detect the content of ROS in hippocampal neurons,Western blot was used to detect the expressions of Drp1,p-Drp1,Mfn2,Opa1,HK1,PKM2,GLUT1 and LDHA,the contents of serum IL-1β,IL-6 and TNF-α were detected by ELISA.Results Compared with the sham-operation group,rats in the model group had a prolonged escape latency(P<0.01)and a reduced number of crossing platforms(P<0.01);neuronal gaps in the CA1 region of the hippocampus were enlarged,with irregular cell morphology and blurred borders,neuronal consolidation,lysis and fragmentation of Nissl bodies and reduced number of Nissl bodies,swelling and deformation of mitochondria,disorganization of the cristae,and disruption of the bilayer membrane structure;the content of ROS in CA1 region of the hippocampus was elevated,the protein expressions of Mfn2 and Opa1 significantly decreased(P<0.01),the expressions of p-Drp1,HK1,PKM2,GLUT1,LDHA proteins significantly increased(P<0.01),and serum contents of IL-1β,IL-6 and TNF-α significantly increased(P<0.01).Compared with the model group,the escape latency was significantly shortened in Ditan Yizhi Decoction groups,positive drug group and inhibitor group(P<0.01),and the number of crossing platforms increased(P<0.05,P<0.01);the number of neurons in the hippocampal CA1 region increased,with normal morphology,orderly arrangement,abundant Nissl bodies,recovered mitochondrial morphology,and decreased rupture;the ROS content in hippocampal CA1 region decreased(P<0.01),while the expressions of Mfn2 and Opa1 proteins increased(P<0.01),the expressions of p-Drp1,HK1,PKM2,GLUT1 and LDHA proteins decreased(P<0.01),and the serum contents of IL-1β,IL-6 and TNF-α decreased(P<0.05).Conclusion Ditan Yizhi Decoction can improve cognitive impairment and neuronal morphology in VaD rats,and the mechanism maybe related to regulation of mitochondrial dynamics through the ROS/Drp1 axis,attenuating glycometabolic disorders,and reducing inflammatory response.

Objective To detect the content of rheumatoid factor(RF)after RF-CIC dissociation using serum circulating immune complexes(CIC)dissociation technology and evaluate its diagnostic and clinical value in rheumatic arthritis(RA).Methods 55 RA patients diagnosed and treated in the 903rd Hospital of the People's Liberation Army from January 2024 to December 2024 were selected as the RA disease group,and 20 healthy individuals were selected as the control group.In addition,57 non RA pa-tients with symptoms resembling RA[patients with systemic lupus erythematosus(SLE),gout,ankylosing spondylitis(AS),osteo-arthritis,etc)]as the non RA disease group.Using CIC dissociation technology,RF content after RF-CIC dissociation was detect-ed in the serum of all three groups of study subjects,and C-reactive protein(CRP)and RF levels in all subjects were detected using a biochemical analyzer.Analyzed and compared the differences in the positive rate and levels of RF-CIC among three groups object of study.In addition,analyze and compare the correlation between RF-CIC and inflammatory index CRP.Results The positive rates of RF-CIC in the serum of RA disease group,non RA disease group,and control group were 87.27%(48/55),10.53%(6/57)and 0.0%(0/20),respectively,and the differences between the three groups was statistically significant(χ2=84.520,P<0.05).Further subgroup analysis showed that the RF-CIC positivity rate in the RF negative subgroup of RA disease patients[61.11%(11/18)]higher than that in the non RA disease group[1.92%(1/52)]and the control group[0%(0/20)],and the differ-ences were statistically significant(χ2=44.493,21.671,all P<0.05).The RF-CIC positivity rate was higher in RF positive pa-tients than in RF negative patients in the RA disease group(100%vs 61.11%),and the difference was statistically significant(χ2=16.487,P<0.05).The RF-CIC content in the serum of RF positive patients in the RA disease group was higher than that of RF negative patients[16.35(10.53,26.49)vs 3.57(2.53,3.89)],and the difference was statistically significant(Z=-4.243,P<0.05).Correlation analysis showed that the levels of CRP and RF in the serum of RA patients were positively correlated with the levels of RF-CIC(r=0.490,0.970,all P<0.05).Conclusion RF-CIC demonstrates high positivity even in RF-negative RA patients,and their levels correlate with CRP.RF-CIC shows potential as a serological indicator for early diagnosis and disease activity assess-ment in RA.

In order to realize the quantitative assessment of muscle strength in hand function rehabilitation and then formulate scientific and effective rehabilitation training strategies, this paper constructs a multi-scale convolutional neural network (MSCNN) - convolutional block attention module (CBAM) - bidirectional long short-term memory network (BiLSTM) muscle strength prediction model to fully explore the spatial and temporal features of the data and simultaneously suppress useless features, and finally achieve the improvement of the accuracy of the muscle strength prediction model. To verify the effectiveness of the model proposed in this paper, the model in this paper is compared with traditional models such as support vector machine (SVM), random forest (RF), convolutional neural network (CNN), CNN - squeeze excitation network (SENet), MSCNN-CBAM and MSCNN-BiLSTM, and the effect of muscle strength prediction by each model is investigated when the hand force application changes from 40% of the maximum voluntary contraction force (MVC) to 60% of the MVC. The research results show that as the hand force application increases, the effect of the muscle strength prediction model becomes worse. Then the ablation experiment is used to analyze the influence degree of each module on the muscle strength prediction result, and it is found that the CBAM module plays a key role in the model. Therefore, by using the model in this article, the accuracy of muscle strength prediction can be effectively improved, and the characteristics and laws of hand muscle activities can be deeply understood, providing assistance for further exploring the mechanism of hand functions .
Neural Networks, Computer ; Humans ; Hand Strength/physiology* ; Support Vector Machine ; Muscle Strength/physiology* ; Hand/physiology* ; Convolutional Neural Networks

ObjectiveTo understand the development stages and use of policy tools of general practitioner policies in China since it was first proposed, to summarize the experience and explore the shortcomings, so as to provide references for the adjustment and optimization of China’s general practitioner policies. MethodsContent analysis and mathematical statistics analysis were used to conduct a quantitative research on 111 policy documents with 422 policy items involving general practitioners at the national level from 1997 to 2023, through a three-dimensional analysis framework integrating policy tools, human capital process and policy development stages. ResultsCapacity‑building policy tools were most frequently used in general practitioner policies, and the policy tools gradually shifted from mandate to inducement. The general practitioner policies paid less attention to the career selection link, but paid full attention to every segment of human capital links, with a comprehensive application of policy tools observed in the integrated development stage, despite the existence of unbalanced internal distribution. ConclusionIt is suggested to promote the use of incentive policy tools and to explore multiple approaches based on incentive theory; pay attention to the career selection link for guiding the employment of general practitioners; take the appropriateness between the policy tools and human capital process into comprehensive consideration, striking a dynamic balance of the internal structure of general practitioner policies.