This paper systematically reviews the core concepts and lines of theoretical inheritance of major spleen-stomach theories in traditional Chinese medicine （TCM）， including spleen deficiency theory， spleen-stomach damp-heat theory， and liver-spleen disharmony theory. It is found that these theories have all undergone a developmental trajectory characterized by classical foundation， refinement of therapeutic methods， systematization of pathogenesis， and modern innovation. The evolution of spleen-stomach theory has achieved a shift from a singular focus on tonifying the spleen to regulating dynamic middle-jiao （焦） balance， and from localized spleen-stomach regulation to the circular movement of qi involving all five zang organs. In terms of modern disease-syndrome integrative research， spleen deficiency syndrome is shown to be closely associated with impairment of the gastrointestinal mucosal barrier， metabolic disorders， and gene polymorphisms related to Helicobacter pylori-associated gastric diseases. Spleen-stomach damp-heat syndrome is closely linked to hyperactive energy metabolism， inflammatory cytokines， and abnormal expression of aquaporins. Liver-spleen disharmony syndrome is mainly associated with dysregulation of the brain-gut axis and microbiota-related metabolic disorders. It is proposed that future research on spleen-stomach diseases and syndromes should further elucidate their potential multidimensional differential biological characteristics， thereby promoting the modernization of the TCM discipline of spleen-stomach studies.

ObjectiveTo investigate the relationship between the composition of traditional Chinese medicine (TCM) and depression/anxiety/sleep disturbances (D/A/S) in patients with chronic pain.MethodsThis cross-sectional study was conducted at 13 tertiary hospitals across China, enrolling patients who experienced chronic pain between November 2023 and May 2024. The Patient Health Questionnaire-9, Generalized Anxiety Disorder-7, Pittsburgh Sleep Quality Index, and TCM constitution categories were used to assess the patients. The association between the TCM constitution and the D/A/S ratio was analyzed using multivariable logistic regression.ResultsA total of 1107 patients (63.2% women) were analyzed. Compared with those with a balanced constitution, patients who had qi-deficiency and yin-deficiency were at a higher risk of depression. Qi-deficiency and yin-deficiency were associated with anxiety. Sleep disturbances were common in patients with qi-deficiency constitution (odds ratio [OR]: 2.32, 95% confidence interval [CI]: 1.42–3.81), yang-deficiency constitution (OR: 1.94, 95% CI: 1.26–2.98), yin-deficiency constitution (OR: 2.03, 95% CI: 1.24–3.32), blood stasis constitution (OR: 2.07, 95% CI: 1.01–4.22), and qi-stagnation constitution (OR: 2.66, 95% CI: 1.35–5.25).ConclusionIn patients with chronic pain, specific TCM constitutions may be associated with D/A/S. Further longitudinal studies are needed to clarify the potential causal relationships between TCM constitution types and these conditions.

OBJECTIVE: To explore the potential role of mRNA m7G modification in the pathogenesis of human adenocarcinoma of esophagogastric junction (AEG). METHODS: Pathological tissue specimens from four AEG patients who underwent surgical treatment at the People's Hospital Affiliated to Jiangsu University between 2018 and 2019 were selected. Tumor tissues and adjacent normal tissues were collected from these patients. RNA was extracted from both tissue types and subjected to m7G methylated RNA immunoprecipitation sequencing (m7G-MeRIP-seq) to analyze the patterns of m7G modification, the characteristics of differential m7G modification sites, the differentially expressed mRNA, and the correlation between m7G modification and mRNA expression levels. Differential m7G-modified genes (MSH6, BRCA1, and SOX9) were further validated using methylated RNA immunoprecipitation quantitative PCR (MeRIP-qPCR), while the expression of METTL1 and WDR4 genes was examined by real-time quantitative PCR (RT-qPCR). This study was approved by the Medical Ethics Committee of the People's Hospital Affiliated to Jiangsu University (Ethics No. 20150083). RESULTS: m7G-MeRIP-seq analysis revealed that m7G modifications in both AEG and adjacent normal tissues were predominantly located in the GC-rich region surrounding the internal start codon of mRNA. Differential m7G modification sites between the two groups were closely associated with cancer-related genes. mRNA library analysis showed that differentially expressed mRNA were predominantly upregulated in AEG tissues and downregulated in adjacent normal tissues. Cross-analysis indicated that genes with hypermethylation tended to exhibit upregulated expression, while genes with hypomethylation were typically downregulated in AEG tissues. MeRIP-qPCR validation confirmed that the mRNA expression of MSH6, BRCA1, and SOX9 were significantly upregulated in AEG tissues compared to adjacent normal tissues (AEG vs. normal, P < 0.05). RT-qPCR results demonstrated that the mRNA expression levels of METTL1 and WDR4 were also upregulated in AEG tissues (AEG vs. normal, P < 0.000 5). CONCLUSION These findings suggest that mRNA m7G modification plays a significant role in the development of AEG. Furthermore, proteins as METTL1 and WDR4 may facilitate AEG progression by regulating mRNA m7G modification. These results provide valuable insights into the molecular mechanisms underlying AEG and may inform future therapeutic strategies for this malignancy.
Humans ; RNA, Messenger/metabolism* ; Adenocarcinoma/pathology* ; Esophagogastric Junction/metabolism* ; Esophageal Neoplasms/metabolism* ; Gene Expression Regulation, Neoplastic ; Female ; Male ; Middle Aged ; DNA Methylation ; Methyltransferases/metabolism* ; Stomach Neoplasms/genetics*

Objective To observe the effects of Chaihu Longgu Muli Decoction on oxidative stress in hippocampus and hypothalamic-pituitary-adrenal(HPA)axis hormone levels of rats with generalized anxiety disorder(GAD);To explore its mechanism of alleviating anxiety in GAD rats.Methods A total of 72 Wistar rats were randomly divided into blank group(12 rats)and modeling group(60 rats).Chronic restraint stress was used to establish the GAD model.The successfully modeled rats were randomly divided into model group,diazepam group(1 mg/kg)and TCM low-,medium-and high-dosage groups(Chaihu Longgu Muli Decoction 6,12,24 g/kg).The administration groups were given corresponding drugs by gavage for 14 consecutive days.Open field experiment and elevated cross maze experiment were used to observe indexes in rat behavior,HE staining was used to observe the morphology of hippocampal tissue,ELISA was used to detect the contents of serum corticotropin releasing hormone(CRH),adrenocorticotropic hormone(ACTH)and corticosterone(CORT),biochemical kits were used to detect the contents of malondialdehyde(MDA),catalase(CAT)and superoxide dismutase(SOD)in hippocampal tissue,RT-PCR was used to detect the mRNA expressions of MEK,ERK,Bax and Bcl-2 in hippocampal tissue,Western blot was used to detect the protein expressions of MEK,p-MEK,ERK,p-ERK,Bax and Bcl-2 in hippocampal tissue.Results Compared with the blank group,the rats in the model group had a significantly slower growth rate of body msss,loose stool,dark hair,significant hair removal and extreme agitation;the activity time and distance in the central area were significantly reduced(P<0.01),the percentage of time,frequency and distance of entering the open arm were significantly reduced(P<0.01);with structural degeneration of hippocampal neurons,changes in cell morphology,disordered arrangement and edema of surrounding tissues;the contents of serum CRH,ACTH and CORT significantly increased(P<0.01);the MDA content in hippocampal tissue significantly increased(P<0.01),CAT and SOD contents significantly decreased(P<0.01),p-MEK and p-ERK protein expressions significantly decreased(P<0.01),Bax mRNA and protein expressions significantly increased(P<0.01),Bcl-2 mRNA and protein expressions significantly decreased(P<0.01).Compared with the model group,the body mass growth rate of diazepam group and each dosage of TCM group increased,and the symptoms of loose stool,hair removal,anxiety and huddling were improved;the activity time and distance in the central area of rats in the diazepam group and TCM medium-and high-dosage groups significantly increased(P<0.01),and the percentage of time,frequency and distance of entering the open arm significantly increased(P<0.05,P<0.01);the contents of serum CRH,ACTH and CORT significantly decreased(P<0.05,P<0.01);the MDA content in the hippocampal tissue in diazepam group and TCM high-dosage group significantly decreased(P<0.01),while the CAT and SOD contents significantly increased(P<0.05,P<0.01),the protein expressions of p-MEK and p-ERK in hippocampal tissue in diazepam group and TCM medium-and high-dosage group significantly decreased(P<0.01),Bax mRNA and protein expressions were significantly decreased(P<0.01),and Bcl-2 mRNA and protein expressions significantly increased(P<0.05,P<0.01).There was no statistical significance in the expressions of MEK,ERK mRNA and protein among the groups(P>0.05).Conclusion Chaihu Longgu Muli Decoction can alleviate anxiety in GAD rats by inhibiting HPA axis hormone release and MEK/ERK signaling pathway activation,improving neuroendocrine levels,and reducing hippocampal oxidative stress response.

Objective:To analyze the drug-resistance pattern and molecular epidemiological characteristics of Enterobacteriales carrying the blaNDM gene in Lishui, aiming to guide clinical anti-infection treatment. Methods:Non-duplicate blaNDM-carrying Enterobacteriales, isolated from Lishui Central Hospital, were collected and identified by VITEK MS. The minimal inhibitory concentrations (MICs) were detected by the broth microdilution method. The ST types of the strains were determined by multilocus sequence typing (MLST). Plasmid types were identified by transformation or conjugation experiments and replication initiator amplification experiments. The transposon structures were detected by PCR amplification. Finally, the epidemic regularity of blaNDM gene in Lishui was analyzed from three levels: clonal group, plasmid, and mobile genetic elements. Results:A total of 109 blaNDM-positive strains were collected. Among them, 60 strains carried the blaNDM-1 gene and 49 strains carried the blaNDM-5 gene. The 109 strains showed 100% resistance to ceftazidime and cefotaxime. The resistance rates to peracillin-tazobactam and imipenem were higher than 80%. Strains carrying the blaNDM-5 gene were more resistant to meropenem than those carrying blaNDM-1 gene( P<0.05). A total of 68 STs were detected from 109 strains, and IncX3, IncFⅡγ, IncA/C and IncT/R plasmids were detected, and 90.83% of the blaNDM genes were located in the IncX3 plasmid. Twelve types of blaNDM gene surrounding structures existed, and they all carried the highly conserved blaNDM- bleMBL- trpF gene sequence. Conclusions:The blaNDM gene has diverse transmission modes in Lishui. The IncX3 plasmid is the main factor mediating its transfer, and all strains carry highly conserved blaNDM- bleMBL- trpF gene sequence.

Objective:To explore the potential role of mRNA m7G modification in the pathogenesis of human adenocarcinoma of esophagogastric junction (AEG).Methods:Pathological tissue specimens from four AEG patients who underwent surgical treatment at the People′s Hospital Affiliated to Jiangsu University between 2018 and 2019 were selected. Tumor tissues and adjacent normal tissues were collected from these patients. RNA was extracted from both tissue types and subjected to m7G methylated RNA immunoprecipitation sequencing (m7G-MeRIP-seq) to analyze the patterns of m7G modification, the characteristics of differential m7G modification sites, the differentially expressed mRNA, and the correlation between m7G modification and mRNA expression levels. Differential m7G-modified genes ( MSH6, BRCA1, and SOX9) were further validated using methylated RNA immunoprecipitation quantitative PCR (MeRIP-qPCR), while the expression of METTL1 and WDR4 genes was examined by real-time quantitative PCR (RT-qPCR). This study was approved by the Medical Ethics Committee of the People′s Hospital Affiliated to Jiangsu University (Ethics No. 20150083). Results:① m7G-MeRIP-seq analysis revealed that m7G modifications in both AEG and adjacent normal tissues were predominantly located in the GC-rich region surrounding the internal start codon of mRNA. Differential m7G modification sites between the two groups were closely associated with cancer-related genes. ② mRNA library analysis showed that differentially expressed mRNA were predominantly upregulated in AEG tissues and downregulated in adjacent normal tissues. ③ Cross-analysis indicated that genes with hypermethylation tended to exhibit upregulated expression, while genes with hypomethylation were typically downregulated in AEG tissues. ④ MeRIP-qPCR validation confirmed that the mRNA expression of MSH6, BRCA1, and SOX9 were significantly upregulated in AEG tissues compared to adjacent normal tissues (AEG vs. normal, P<0.05). ⑤ RT-qPCR results demonstrated that the mRNA expression levels of METTL1 and WDR4 were also upregulated in AEG tissues (AEG vs. normal, P<0.000 5). Conclusion:These findings suggest that mRNA m7G modification plays a significant role in the development of AEG. Furthermore, proteins as METTL1 and WDR4 may facilitate AEG progression by regulating mRNA m7G modification. These results provide valuable insights into the molecular mechanisms underlying AEG and may inform future therapeutic strategies for this malignancy.

Objective:To analyze the genetic characteristics of clinical manifestations in children with KBG syndrome due to microdeletions.Methods:A retrospective case summary was conducted. Four children diagnosed with KBG syndrome due to 16q24.3 microdeletion at Children′s Hospital of Zhengzhou University from July 2021 to April 2024 were enrolled.Their clinical manifestations, biochemical parameters, imaging data, whole-exome sequencing results, treatments and follow-up outcomes were reviewed.Results:The cohort included two males and two females (diagnosed at 81, 18, 26, and 56 months of age, respectively), from four unrelated families. All patients exhibited peculiar facial features (Cupid′s bowed-shaped lips, prominent ears, thick eyebrows), skeletal abnormalities (brachydactyly, abnormal ribs, short stature, etc.), ocular anomalies (astigmatism, strabismus, amblyopia, etc.), intrauterine growth restriction, and developmental retardation. Case 2, 3, 4 had cranial imaging abnormalities, including thin anterior pituitary lobes with pineal cyst, left ventricular cyst, and abnormal pituitary stalk or lateral ventricles with sinusitis, respectively. Two children had intellectual disability, two had congenital heart disease, and one had delayed bone age and hair abnormalities. Whole exome genomic sequencing confirmed 16q24.3 microdeletions encompassing ANKRD11 gene in all four cases. Two children treated with recombinant human growth hormone achieved height increments of 1.5 s and 0.4 s, respectively. Conclusions:Typical features of 16q24.3 microdeletion-induced KBG syndrome include peculiar facial features, macrodontia, skeletal anomalies, neurological abnormalities, and ocular defects. Genetic testing is essential for definitive diagnosis. The treatment of KBG syndrome requires early diagnosis and multidisciplinary collaboration to implement individualized treatment for multisystem symptoms.

Objective:To compare the differences among four routine lipid testing systems in detecting high triglyceride (TG) serum samples and evaluate the accuracy and consistency of the four homogeneous low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) reagents using vertical auto profile (VAP) as the reference method.Methods:A retrospective study was conducted on 249 serum samples with elevated TG levels collected from the Department of Laboratory Medicine at the Second Xiangya Hospital of Central South University between January and October 2024. TG, total cholesterol (TC), LDL-C, and HDL-C were measured using four homogeneous detection systems: Beckman Coulter (USA), Wako Pure Chemical Industries (Japan), Mindray (China), and Roche Diagnostics (Germany). VAP was used to analyze lipoprotein subfractions, including very-low-density lipoprotein cholesterol (VLDL-C), intermediate-density lipoprotein cholesterol (IDL-C), LDL-C, lipoprotein(a) cholesterol [Lp(a)-C], and HDL-C. The mean coefficient of variation ( CV) across the four systems was calculated for each parameter. Pearson correlation and ordinal logistic regression (OLR) were used to assess correlations between the four HDL-C/LDL-C systems and VAP. Bland-Altman plots were generated to evaluate biases, and deviations were calculated. For parameters with significant deviations, multivariate linear regression and standardized coefficients were used to analyze correlations between biases and lipoprotein subfractions. Based on the Chinese Guidelines for Lipid Management (2023), LDL-C and non-HDL-C treatment goals were categorized into five risk levels (ultra-high, high, moderate, high-risk, and low-risk). VAP results defined LDL-C/non-HDL-C intervals, and the four systems′ concordance in risk classification was evaluated. Samples were grouped into A, B, C, D ( n=63, 62, 62, 62) by TG concentration, and ANOVA, chi-square, and Fisher exact tests assessed intergroup differences. Results:The mean CVs across systems for TG, TC, LDL-C, HDL-C, and non-HDL-C were 2.98%, 1.76%, 18.10%, 5.60%, 2.58%, respectively. Pearson correlations between LDL-C results (Beckman, Wako, Mindray, Roche) and VAP were 0.889, 0.854, 0.899, and 0.973; mean relative deviations were 54.8%, 41.0%, 49.3%, and 3.6%; classification accuracies were 6.0% (15/249), 21.3% (53/249), 9.2% (23/249), and 76.7% (191/249). HDL-C deviations were 18.7%, 15.1%, 11.1%, and 8.7%, with correlations ( r) of 0.883, 0.911, 0.959, and 0.950 (all P<0.001). LDL-C means showed no intergroup differences (A-D), but CV increased with TG levels ( P<0.001). HDL-C means and CVs showed no significant intergroup differences. Beckman, Wako, and Mindray LDL-C results exhibited significant positive biases correlated with TG and VLDL-C (multivariate regression; P<0.05); VLDL-C had the strongest influence (standardized coefficients: 0.820, 0.394, 0.813; P<0.001). Non-HDL-C classifications matched VAP in 92.4% (Beckman), 85.9% (Wako), 94.0% (Mindray), and 93.2% (Roche), with no intergroup differences. Conclusion:For high-TG sera, Beckman, Wako, and Mindray LDL-C exhibited significant positive biases correlated with TG and VLDL-C, while Roche LDL-C showed minimal deviation. TG, TC, HDL-C, and non-HDL-C results showed minimal variation across the four systems. All systems demonstrated comparable accuracy for non-HDL-C compared to VAP. The non-HDL-C measured by the four detection systems demonstrates high accuracy and consistency in atherosclerotic cardiovascular disease risk stratification and lipid-lowering goal assessment, and it is unaffected by TG levels.

Bio-mineralization has emerged as a promising strategy to enhance vaccine immunogenicity. This study optimized the calcium phosphate (CaP) mineralization process of foot-and-mouth disease virus-like particles (FMD VLPs) to achieve high mineralization efficiency and scalability. Key parameters, including concentrations of Ca²⁺, HPO₄^2-, NaCl, and VLPs, as well as stirring speed, were systematically optimized. Stability of the scaled-up reaction system and immunogenicity of the mineralized vaccine were evaluated. Optimal conditions [25.50 mmol/L Ca(NO₃)₂, 15 mmol/L Na₂HPO₄, 300 mmol/L NaCl, 0.75 mg/mL VLPs, and 1 500 r/min] yielded CaP-mineralized VLPs (VLPs-CaP) with high mineralization efficiency, uniform morphology, and a favorable particle size. Scaling up the reaction by 25 folds maintained consistent mineralization efficiency and particle characteristics. Immunization in mice demonstrated that VLPs-CaP induced higher titers of specific antibodies and neutralizing antibodies than unmineralized VLPs (P < 0.05). Higher IgG2a/IgG1 ratio and enhanced IFN-γ secretion (P < 0.05) further indicated robust cellular immune responses. We establish a stable and scalable protocol for VLPs-CaP, providing a theoretical and technical foundation for developing high-efficacy VLPs-CaP vaccines.
Vaccines, Virus-Like Particle/immunology* ; Immunogenicity, Vaccine ; Calcium Phosphates/chemistry* ; Foot-and-Mouth Disease Virus ; Biomineralization ; Particle Size ; Animals ; Mice ; Antibodies, Neutralizing/blood* ; Antibodies, Viral/blood* ; Immunity, Cellular

Vaccination is a crucial strategy for the prevention and control of infectious diseases. Virus-like particles (VLPs), composed of structural proteins, have garnered significant attention as a novel type of vaccine due to their excellent safety and immunogenicity. However, similar to most vaccine antigens, VLPs exhibit insufficient thermal stability, which not only restricts the widespread application of vaccines but also increases the risk of vaccine inactivation. This study aims to enhance the stability and shelf life of VLPs derived from type A foot-and-mouth disease virus (FMDV) by employing vacuum freeze-drying technology. The optimal lyoprotectant formulation was determined through single-factor and combinatorial screening. Subsequently, the correlation between the immunogenicity of the freeze-dried vaccine and the content of FMDV VLPs was evaluated via a mouse model. The stability of FMDV VLPs before and after freeze-drying was further assessed by storing them at 4, 25, and 37 ℃ for varying time periods. Results indicated that the lyoprotectant formulation No.1, composed of 7.5% trehalose, 0.1% Tween 80, 50 mmol/L glycine, 1% sodium glutamate, and 3% polyvinylpyrrolidone (PVP), effectively preserved the content of FMDV VLPs during the vacuum freeze-drying process. The immunization trial in mice revealed that the levels of specific antibodies, immunoglobulin G1 (IgG1), interleukin-4 (IL-4), and neutralizing antibodies induced by freeze-dried FMDV VLPs were comparable to those induced by non-freeze-dried FMDV VLPs. The heat treatment results showed that the storage periods of freeze-dried FMDV VLPs at 4, 25, and 37 ℃ were significantly longer than those of non-freeze-dried FMDV VLPs. In conclusion, the selected lyoprotectant formulation effectively improved the stability of FMDV VLPs vaccines. This study provides valuable insights for enhancing the stability of novel subunit vaccines.
Freeze Drying/methods* ; Animals ; Foot-and-Mouth Disease Virus/immunology* ; Mice ; Vaccines, Virus-Like Particle/chemistry* ; Foot-and-Mouth Disease/immunology* ; Vacuum ; Drug Stability ; Mice, Inbred BALB C ; Viral Vaccines/immunology*