Objective To investigate the characteristics of Chinese medicine syndromes and the possible metabolic substance basis of spontaneously hypertensive rat(SHR).Methods 10-week-old SPF SHR and WKY of the same strain were divided into SHR group and WKY group with 8 rats in each group.The general state,temperament,peripheral vascular filling,tongue characteristics,diet,water intake,urine and feces volume and characteristics,blood pressure,heart rate,respiratory rate,pain threshold,and open field behavior of SHR rats were observed and tested comprehensively to identify the possible syndrome types of Chinese medicine.At the same time,liquid chromatography tandem mass spectrometry was used to analyze non-targeted serum metabolites to preliminarily reveal the material basis of blood pressure elevation and Chinese medicine syndrome manifestations.Results Compared with WKY group,the scores of dark yellow hair color,irritable degree and peripheral capillary filling were higher in SHR group(P＜0.0001).Red tongue color,dry tongue,little body fluid;24 h diet and water intake,urine volume and fecal volume were less(P＜0.05),fecal water content was lower(P＜0.001);systolic blood pressure(SBP),diastolic blood pressure(DBP),mean arterial pressure(MAP),heart rate(HR)and respiratory rate(RR)were significantly higher(P＜0.05);Lower pain threshold(P＜0.0001);The open field experiment shows that the moving distance and residence time of the edge are longer(P＜0.001).Serum non-targeted metabolomics result showed that,compared with WKY group,the SHR group had 114 metabolites with significant differences(P＜0.05).These differential metabolites were mainly lipids and lipid-like molecules(40.35％),organic acids and derivatives(22.8％),and organoheterocyclic compounds(15.79％).A total of 25 metabolic pathways were identified by KEGG enrichment analysis.Further differential abundance analysis showed that 16 pathways were activated,only 4 pathways were inhibited,and 5 pathways were not significantly changed.The glutamatergic synapse and GABAergic synapse were activated,while the serotonergic synapse was inhibited.Conclusions The symptoms of SHR include impatience and irritability,peripheral vascular dilation and collateral circulation formation,bulbar conjunctival congestive swelling,red tongue coloration,a dry tongue,constipation,red-yellow urine of low volume,and a rapid heart rate and high respiratory rate.All these suggest that SHR is a syndrome of hypertension with hyperactivity of liver-yang.The material basis of SHR is not only related to lipid,amino acid,and carbohydrate metabolism disorders,but also may be related to metabolic disorders of glutaminergic,GABAergic,and serotonergic neural pathways.

Objective To investigate the characteristics of Chinese medicine syndromes and the possible metabolic substance basis of spontaneously hypertensive rat(SHR).Methods 10-week-old SPF SHR and WKY of the same strain were divided into SHR group and WKY group with 8 rats in each group.The general state,temperament,peripheral vascular filling,tongue characteristics,diet,water intake,urine and feces volume and characteristics,blood pressure,heart rate,respiratory rate,pain threshold,and open field behavior of SHR rats were observed and tested comprehensively to identify the possible syndrome types of Chinese medicine.At the same time,liquid chromatography tandem mass spectrometry was used to analyze non-targeted serum metabolites to preliminarily reveal the material basis of blood pressure elevation and Chinese medicine syndrome manifestations.Results Compared with WKY group,the scores of dark yellow hair color,irritable degree and peripheral capillary filling were higher in SHR group(P＜0.0001).Red tongue color,dry tongue,little body fluid;24 h diet and water intake,urine volume and fecal volume were less(P＜0.05),fecal water content was lower(P＜0.001);systolic blood pressure(SBP),diastolic blood pressure(DBP),mean arterial pressure(MAP),heart rate(HR)and respiratory rate(RR)were significantly higher(P＜0.05);Lower pain threshold(P＜0.0001);The open field experiment shows that the moving distance and residence time of the edge are longer(P＜0.001).Serum non-targeted metabolomics result showed that,compared with WKY group,the SHR group had 114 metabolites with significant differences(P＜0.05).These differential metabolites were mainly lipids and lipid-like molecules(40.35％),organic acids and derivatives(22.8％),and organoheterocyclic compounds(15.79％).A total of 25 metabolic pathways were identified by KEGG enrichment analysis.Further differential abundance analysis showed that 16 pathways were activated,only 4 pathways were inhibited,and 5 pathways were not significantly changed.The glutamatergic synapse and GABAergic synapse were activated,while the serotonergic synapse was inhibited.Conclusions The symptoms of SHR include impatience and irritability,peripheral vascular dilation and collateral circulation formation,bulbar conjunctival congestive swelling,red tongue coloration,a dry tongue,constipation,red-yellow urine of low volume,and a rapid heart rate and high respiratory rate.All these suggest that SHR is a syndrome of hypertension with hyperactivity of liver-yang.The material basis of SHR is not only related to lipid,amino acid,and carbohydrate metabolism disorders,but also may be related to metabolic disorders of glutaminergic,GABAergic,and serotonergic neural pathways.

This study aimed to elucidate the mechanism of action of metformin (MET) in inhibiting the malignant progression of hepatocellular carcinoma (HCC) by regulating the degradation of aldo-keto reductase family 1 member C3 (AKR1C3). The correlation between the sensitivity of different hepatocellular carcinoma cell lines to MET and their basal expression levels of AKR1C3 was firstly evaluated. MET was found to significantly reduce the level and accelerate the degradation rate of AKR1C3 protein by Western blot. The interaction between MET and AKR1C3 protein was confirmed by cellular thermal shift assay (CETSA). Proteasome inhibitor MG132 and the lysosomal inhibitor chloroquine (CQ) were used to screen the degradation pathway, and confirm, in combination with the HBSS starvation-induced autophagy model, that MET mediated the degradation of AKR1C3 through the autophagy lysosome pathway. Ubiquitylation assay showed that MET specifically enhanced the K63-linked polyubiquitylation modification of AKR1C3. Sequestosome 1 (SQSTM1/p62) knockdown, immunoprecipitation, and immunofluorescence co-localization analyses confirmed that the autophagy receptor p62 plays a key role in mediating MET-induced degradation of AKR1C3. The adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) inhibitor compound C was used to demonstrate that the regulatory effect of MET on AKR1C3 is independent of the classical AMPK signaling pathway. The experimental results showed that metformin promoted the ubiquitination modification of AKR1C3 by targeting AKR1C3, enhanced the binding of AKR1C3 to autophagy receptor p62, then degraded the AKR1C3 protein through selective autophagy-like pathway, and ultimately inhibited the malignant phenotypes of hepatocellular carcinoma cells, which is a regulatory mechanism free of the classical AMPK activation pathway of metformin.

Objective:To explore the clinical characteristics, audiological outcomes, and factors influencing the efficacy of pharmacological treatment in patients with sudden hearing loss associated with large vestibular aqueduct syndrome（LVAS）. Methods:A retrospective analysis was conducted on the clinical data of 77 bilateral LVAS patients（117 ears） hospitalized for sudden hearing loss from January 1, 2009, to December 31, 2023. The inclusion criteria required that patients to be diagnosed according to the Valvassori standard and had received standardized pharmacological treatment. Clinical features, audiological outcomes, and treatment efficacy were analyzed. Statistical methods were employed to identify factors associated with treatment outcomes. Results:The age of the enrolled patients ranged from 4 to 37 years. The age of onset for the initial hearing fluctuation varied between 0 and 24 years, with a mean age of 5.8 years. The male-to-female ratio was approximately balanced（37 males and 40 females）. The proportion of unilateral to bilateral sudden hearing loss was 1.0︰1.2, with unilateral right ear hearing loss being more frequently occurring（64.9%）. Triggering Factors: Triggers included no identifiable factors in 48.1% of cases, a history of head trauma（24.7%）, upper respiratory tract infections（11.7%）, onset following physical fatigue（11.7%）, and less frequently, noise exposure, alcohol consumption, or emotional stress（each 1.3%）. Clinical Symptoms: Hearing loss was the sole symptom in 35.1% of cases. Concurrent symptoms included vertigo in 44.2% and tinnitus in 46.8%. Patients with a disease duration of ≤14 days demonstrated a treatment efficacy rate of 75.0%. Among those who responded to treatment, 93.0% had profound or greater hearing loss prior to therapy, with an average improvement in hearing thresholds of 32 dB HL. In pretreatment, 68.9% of patients exhibited low-frequency air-bone gaps, increasing to 76.1% post-treatment. Additionally, 17.6% of treated ears demonstrated a ≥15 dB HL improvement in low-frequency bone conduction thresholds. In the non-responsive group, 7.3% of ears still showed some improvement in bone conduction thresholds. Statistically significant differences（P<0.05） were observed between the treatment-effective and non-effective groups concerning the age of initial hearing fluctuation, disease duration, and severity of hearing loss at onset. Conclusion:The efficacy of pharmacological treatment for sudden hearing loss in LVAS patients is influenced by the age at onset, duration of the disease, and severity of hearing impairment. Early diagnosis and timely intervention significantly enhance treatment efficacy, particularly in patients with a disease duration of ≤14 days and an initial sudden hearing loss. Patients with severe hearing loss, especially those with profound or greater impairment, exhibit greater sensitivity to treatment. Pharmacological interventions positively impact both air conduction and bone conduction thresholds, with the observed improvement in bone conduction thresholds warranting further investigation.
Humans ; Male ; Retrospective Studies ; Hearing Loss, Sudden/etiology* ; Female ; Vestibular Aqueduct/pathology* ; Adult ; Child ; Adolescent ; Child, Preschool ; Young Adult ; Treatment Outcome

ObjectiveTo observe the effect of Zhengan Xifengtang on blood pressure and fecal microflora of spontaneously hypertensive rats （SHRs）. MethodA total of 75 male SHRs aged nine weeks were randomly divided into SHR group， Benazepril group （1.00 mg·kg^-1·d^-1）， high-dose Zhengan Xifengtang group （34.5 g·kg^-1·d^-1）， medium-dose Zhengan Xifengtang group （17.25 g·kg^-1·d^-1）， and low-dose Zhengan Xifengtang group （8.625 g·kg^-1·d^-1）. A total of 15 male Wistar-Kyoto （WKY） rats aged nine weeks were selected as the normal group. The normal group and SHR group were administrated with an equal volume of distilled water for eight weeks. During the administration， the blood pressure of the rats was measured regularly. After the intervention， fresh feces were collected with a sterile frozen storage tube， and 16S amplicon information was collected and analyzed. Plasma， hippocampus， and ileum of rats were collected for ultra-high performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS） detection. ResultZhengan Xifengtang decreased the systolic blood pressure and diastolic blood pressure of SHRs. Compared with the SHR group， Zhengan Xifengtang decreased the diversity of fecal microflora of SHRs. At the phylum level， Zhengan Xifengtang increased the relative abundance of SHR Verrucomicrobia and Actinobacteria and decreased the relative abundance of Synergistetes， Tenericutes， and Candidatus Saccharibacteria. Compared with the SHR group， Zhengan Xifengtang increased the relative abundance of Blautia wexlerae， Fusicatenibacter saccharivorans， and Akkermansia muciniphila and decreased the relative abundance of Clostridium lavalense， Intestinimonas butyriciproducens， Acetatifactor muris， Alloprevotella rava， and Oscillibacter valericigenes. Spearman correlation analysis showed that the systolic blood pressure of rats was negatively correlated with the relative abundance of Ethanoligenens， Aerococcus， Butyrivibrio， Olsenella， Bifidobacterium， Clostridium XIVb， Allobaculum， and Fusicatenibacter and positively correlated with the relative abundance of Alloprevotella. Zhengan Xifengtang increased the contents of plasma， hippocampal 5-hydroxy tryptamine（5-HT）， and 5-hydroxyindole acetic acid（5-HIAA） in SHRs and decreased the contents of 5-HT and 5-HIAA in the ileum， and the content of 5-HT in the hippocampus was negatively correlated with that in the ileum. ConclusionZhengan Xifengtang can reduce the blood pressure of SHRs， which may be related to reducing the diversity of SHR microflora， regulating the structure of the microflora， increasing the relative abundance of 5-HT and short-chain fatty acids bacteria， and lowering the relative abundance of pathogenic bacteria related to intestinal inflammation.

There is a bidirectional relationship between intestinal flora and bile acids,and the imbalance of intestinal flora-bile acid axis metabolism is closely related to hypertension.Based on classical TCM literature and clinical practice,this article found that"earth deficiency"is the important pathological basis of hypertension,"wood depression"is the initiating factor of hypertension,and"earth deficiency and wood depression"is the key pathogenesis of hypertension.Combined with the research results of modern medicine and molecular biology,it is considered that the imbalance of intestinal flora and abnormal bile acid metabolism are closely related to the"earth deficiency"and"wood depression"of TCM respectively,and the imbalance of intestinal flora-bile acid axis coincides with the"earth deficiency and wood depression"of TCM in the process of hypertension.It is of great theoretical and practical significance to explore the biological connotation of hypertension"earth deficiency and wood depression"from the perspective of intestinal flora-bile acid axis for guiding TCM to prevent and treat hypertension.

In vitro experiments are an indispensable method in the pharmacological research of Chinese materia medica. However， the development of in vitro experiments is hindered by the various sources， complex composition， and unstable physical and chemical properties of Chinese materia medica. Therefore， it has become an urgent problem to develop reasonable methods for in vitro pharmacological research. At present， the commonly used in vitro experimental methods in the pharmacological research of Chinese materia medica include direct administration and indirect administration. Indirect administration methods include drug-containing serum/plasma， drug-containing intestinal absorption solution， and drug-containing liver incubation solution. With the development of computer technology and systems biology， the combination of high performance liquid chromatography-mass spectrometry （HPLC-MS）， omics， network pharmacology， computer/artificial intelligence-assisted drug design and other technologies with in vitro experiments of Chinese materia medica has not only greatly reduced the research cost but also significantly improved the efficiency of pharmacological exploration， effective ingredient screening， and drug discovery. Therefore， the combined methods have been gradually recognized and widely used. This paper reviewed the in vitro experimental methods in pharmacological research of Chinese materia medica and their progress， aiming to provide help for selecting reasonable research methods.

Objective:To explore the predictive value of red blood cell distribution width-to-platelet ratio (RPR) on arteriovenous fistula (AVF) dysfunction in maintenance hemodialysis patients with end-stage renal disease (ESRD).Methods:One hundred and five ESRD patients who underwent maintenance hemodialysis with autologous AVF in Jiangdu People′s Hospital from March 2021 to March 2023 were selected. The red blood cell distribution width (RDW) and platelet (PLT) levels were measured before the start of dialysis, and the RPR was calculated. All patients were followed up for 6 months, and AVF dysfunction was recorded and divided into dysfunction group and non dysfunction group. The levels of RDW, PLT, and RPR before dialysis were compared between the two groups. Univariate and multivariate Logistic regression analyses were used to examine the relationship between RDW, PLT, RPR and AVF dysfunction in ESRD maintenance hemodialysis patients. Receiver operating characteristic (ROC) curves were plotted to obtain the area under the curve (AUC) for RDW analysis the predictive value of PLT and RPR for AVF dysfunction in ESRD maintenance hemodialysis patients.Results:Among the 105 patients, 4 were excluded due to interruption of treatment and transfer to another hospital. Among the 101 patients ultimately enrolled, 19 patients experienced AVF dysfunction (dysfunction group), with an incidence rate of 18.81% (19/101); 80 patients did experience AVF dysfunction (non dysfunction group). C-reactive protein, RDW and RPR in the dysfunction group were higher than those in the non dysfunction group: (7.36 ± 1.92) mg/L vs. (5.90 ± 2.40) mg/L, (17.98 ± 2.40)% vs. (14.96 ± 2.29)%, 0.14 ± 0.03 vs. 0.11 ± 0.02, the proportion of diabetes patients was higher than that in the non dysfunction group, while albumin was lower than that in the non dysfunction group: (33.49 ± 2.78) g/L vs. (35.01 ± 3.02) g/L, with a statistical significant difference ( P<0.05). Multivariate Logistic regression analysis showed that C-reactive protein, RDW, RPR were all associated with AVF dysfunction in ESRD maintenance hemodialysis patients ( P<0.05). ROC curve was drawn, and the results showed that the AUC of RDW and RPR predicting AVF dysfunction in ESRD maintenance hemodialysis patients was greater than 0.7, and the AUC of RPR was higher, 0.840 (95% CI 0.752 to 0.928), with an optimal cutoff value of 0.125, specificity of 78.90%, and sensitivity of 73.20%. Conclusions:RDW and RPR are both associated with AVF dysfunction in ESRD maintenance hemodialysis patients, and may have certain predictive value for AVF dysfunction.

Objective To study the expression of high mobility group box protein 4 (SOX4) and β-catenin in lung tissues of preterm rats with hyperoxia exposure,and to study its significance.Method One-day-old preterm Sprague Dawley rats were randomly assigned into 4 groups:95％ hyperoxia group,70％ hyperoxia group,45％ hyperoxia group and air group.After three days,pathological changes of the lung tissues were observed by hematoxylin eosin staining,the mRNA expressions of SOX4 and β-catenin in lung tissues were detected by semi-quantitative reverse transcription polymerase chain reaction,and the protein expressions of SOX4 and β-catenin in the lung tissues were measured by western blot.Result The structure of lung tissues in air group was normal.45％ hyperoxia group represented mainly effusion and inflammatory cell infiltration,70％ and 95％ hyperoxia group showed acute lung injury characterized by inflammatory cell infiltration,hyperaemia,hemorrhagic change,disorganization and collapse of alveolar.The expressions of SOX4 mRNA and protein were highly elevated in all hyperoxia groups than that in air group (P ＜ 0.01);Compared with 45％ hyperoxia group,the expressions of SOX4 mRNA and protein were higher in 95％ hyperoxia group (P ＜ 0.01).β-catenin mRNA,total β-catenin protein and nuclear β-catenin protein were also highly elevated in hyperoxia groups compared with air group (P ＜ 0.05).Compared with 45％ hyperoxia group,the expressions of β-catenin mRNA and total β-catenin protein were also highly raised in 70％ hyperoxia group and 95％ hyperoxia group (P ＜ 0.05).The expression of nuclear β-catenin protein was higher in 95％ hyperoxia group than that in 45％ hyperoxia group (P ＜ 0.01).Conclusion The mRNA and protein expressions of SOX4 and β-catenin in lung tissues of preterm rats were increased by hyperoxia exposure.This mechanism may take part in hyperoxia-induced preterm rats lung injury.

Objective To investigate the value of anti-human leukocyte antigen (HLA)antibody level detected by Luminex testing in predicting clinical prognosis of renal transplantation recipients. Methods A total of 1 105 patients scheduled to undergo renal transplantation (354 successfully undergoing renal transplantation)in the 181st Hospital of People's Liberation Army from June 2013 to November 2015 were selected. The serum samples were collected from 1 923 cases before and after renal transplantation. The positive rate and fluorescent intensity of anti-HLA antibody were detected by Luminex testing before and after renal transplantation. The renal function of recipients was also evaluated after renal transplantation. Results Prior to renal transplantation,51.0%(546/1 071)of serum samples were positive for anti-HLA antibody,including 26.0%(279/1 071)positive for anti-HLAⅠantibody,24.9%(267/1 071)positive for anti-HLAⅡantibody and 11.4% (122/1 071 )positive for both anti-HLA Ⅰ and anti-HLA Ⅱ antibodies. Among 354 patients undergoing renal transplantation,59 (17%)were positive for anti-HLA antibody after renal transplantation,including 25 (4 newly positive after surgery)positive for anti-HLAⅠantibody,15 (1 newly positive after surgery)positive for anti-HLAⅡantibody and 19 (4 newly positive after surgery)positive for both anti-HLA Ⅰ and anti-HLA Ⅱ antibodies. During subsequent follow-up,13 patients positive for anti-HLAⅠantibody,5 positive for anti-HLAⅡantibody and 1 1 positive for both anti-HLA Ⅰ and anti-HLA Ⅱ antibodies developed transplant kidney dysfunction. All patients newly positive for anti-HLA antibody after renal transplantation presented with transplant kidney dysfunction. Conclusions Luminex testing can perform dynamic detection of the positive rate of anti-HLA antibody,which is important in predicting clinical prognosis of recipients after renal transplantation.