AIM: To identify the primary active components and underlying mechanisms of Yijing Decoction(YJD)in treating early diabetic retinopathy(DR)based on liquid chromatography-mass spectrometry and network pharmacology.METHODS: Active components of YJD were characterized through LC-MS. Components with optimal ADME(absorption, distribution, metabolism, excretion)properties were selected as key bioactive candidates. Network pharmacology approaches were employed to predict YJD-DR therapeutic targets. Protein-protein interaction(PPI)networks, gene ontology(GO)enrichment analysis, and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis were subsequently conducted to predict core targets and networks. Critical targets and pathways were experimentally validated through Western blot.RESULTS: Ten core therapeutic targets were identified, including TNF, Alb, EGFR, STAT3, PTGS2, ESR1, PPAR, MMP9, TLR4, and MAPK. YJD was related to cancer-related signaling, fluid shear stress and atherosclerosis, and neurodegenerative diseases, encompassing key biological processes such as inflammatory response regulation, programmed cell death activation, and enhanced cell migration. Furthermore, Western blot analysis confirmed that YJD significantly inhibited high glucose-induced phosphorylation of STAT3(P-STAT3/STAT3)and ERK(P-ERK/ERK)in rat retinal microvascular endothelial cells.CONCLUSION: This study revealed YJD's pharmacodynamical basis and its multi-component, multi-target, and multi-paths pharmacology. YJD exerts therapeutic effects on DR by coordinately regulating critical signaling pathways and alleviating intraocular inflammation, thus preserving retinal vascular endothelial cells, maintaining blood-retinal barrier integrity, and facilitating retinal neurovascular repair.

AIM: To predict diabetic retinopathy(DR)progression through macular layer thickness in diabetic patients detected by optical coherence tomography(OCT).METHODS: Retrospective study. The clinical data of 100 cases(200 eyes)of diabetic patients admitted to our hospital from January 2023 to September 2024 were collected. According to the international clinical DR classification, they were divided into the non-diabetic retinopathy(NDR)group with 32 cases(64 eyes), the non-proliferative diabetic retinopathy(NPDR)group with 38 cases(76 eyes), and the proliferative diabetic retinopathy(PDR)group with 30 cases(60 eyes). At the same time, 49 cases(98 eyes)of healthy controls whose age and gender were matched with those of the diabetic patients were collected as the normal group. All patients underwent OCT examination. The thickness changes of the retinal nerve fiber layer(RNFL), ganglion cell layer(GCL), inner plexiform layer(IPL), outer nuclear layer(ONL), photoreceptor cell layer and total retinal thickness(RT)in the subregions of the macular area were compared among the groups. The Eta coefficient was used to analyze the correlation between them and the severity of DR.RESULTS: The thickness of RNFL, GCL, IPL, ONL and photoreceptor cell layer in each sub-region and the average of macular area in the PDR group was significantly lower than that in the NDR and normal groups, while the average RT thickness was significantly higher than that in the NPDR, NDR and normal groups(all P<0.05). The thickness of RNFL(central area, upper inner and outer rings and lower inner and outer rings and average), GCL(upper inner and outer rings and lower inner and outer rings and average), IPL(upper inner ring), ONL(central, upper inner ring and lower inner ring)and photoreceptor cell layer(upper inner and outer rings and lower inner and outer rings and average)in macular area of the PDR group was significantly thicker than that in the NPDR group(all P<0.05). The thickness of RNFL, GCL, IPL, ONL and photoreceptor cell layer in each sub-region and the average of macular area in the NPDR group was significantly lower than that in the NDR and normal groups, while the average RT thickness was significantly thicker than that in the NDR and normal groups(all P<0.05). There was no statistically significant difference in the above indicators between the NDR group and the normal group(all P>0.05). The severity of DR was significantly correlated with the average thickness of RNFL, GCL, IPL, ONL, photoreceptor cell layer and RT in macular area(all P<0.001).CONCLUSION: OCT measurement of the thickness of RNFL, GCL, IPL, ONL, photoreceptor cell layer and RT in the macular area in the diabetic patients can evaluate the progression of DR.

A high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) method was established for simultaneous determination of seven characteristic components from the active fraction of Alpiniae Officinarum Rhizoma in rat plasma, including galangin, kaempferol, kaempferide, pinocembrin, 1,7-diphenyl-4-en-3-heptanone, 5-hydroxy-7-(4-hydroxy-3-methoxyphenyl)-1-phenyl-3-heptanone(DHPA), and 7-(4-hydroxy-3-methoxyphenyl)-1-phenyl-4-en-3-heptanone(DPHB). The new developed HPLC-MS/MS method was applied to study the pharmacokinetics of the 7 characteristic components in rats with Helicobacter pylori gastritis. A Waters Sunfire C_(18) column(2.1 mm×150 mm, 3.5 μm) was used. The acetonitrile-aqueous solution(containing 0.1% formic acid) was adopted as the mobile phase for gradient elution. Seven components and internal standard(chlorogenic acid) were separated within 12 min. Mass spectrometric detection was performed in multiple reaction monitoring(MRM) mode using electrospray ionization(ESI) source with fast switching between positive and negative ions. The method was verified by specificity, linearity, precision, accuracy, recovery, matrix effect, and stability and met the requirements of pharmacokinetic study on the 7 components in rat plasma. Pharmacokinetic results showed that the average peak time(T_(max)) of the 7 components was 0.31-2.19 h, their elimination half-life(t_(1/2)) was 5.26-16.65 h, and the average residence time(MRT) was 6.29-31.03 h after the oral administration of the active fraction of Alpiniae Officinarum Rhizoma to rats with H. pylori gastritis. The plasma exposure levels of galangin and DHPA were higher than those of the other components. The concentration-time curves of four detected flavonoids showed obvious double peaks. This study elucidated the pharmacokinetic characteristics of 7 characteristic components from the active fraction of Alpiniae Officinarum Rhizoma in rats with H. pylori gastritis, providing a scientific basis for the identification of the pharmacodynamic substances of Alpiniae Officinarum Rhizoma for treatment of H. pylori gastritis and the clinical application of Alpiniae Officinarum Rhizoma in the prevention and treatment of H. pylori gastritis.
Animals ; Rats ; Chromatography, High Pressure Liquid/methods* ; Tandem Mass Spectrometry/methods* ; Drugs, Chinese Herbal/administration & dosage* ; Male ; Helicobacter pylori/drug effects* ; Alpinia/chemistry* ; Rats, Sprague-Dawley ; Gastritis/metabolism* ; Helicobacter Infections/metabolism* ; Flavonoids/blood* ; Rhizome/chemistry* ; Liquid Chromatography-Mass Spectrometry

OBJECTIVES: To evaluate the effectiveness of single-balloon and double-balloon enteroscopy in diagnosing pediatric small bowel diseases and assess the diagnostic efficacy of computed tomography enterography (CTE) for small bowel diseases using enteroscopy as the reference standard. METHODS: Clinical data from 576 children who underwent enteroscopy at Hunan Children's Hospital between January 2017 and December 2023 were retrospectively collected. The children were categorized based on enteroscopy type into the single-balloon enteroscopy (SBE) group (n=457) and double-balloon enteroscopy (DBE) group (n=119), and the clinical data were compared between the two groups. The sensitivity and specificity of CTE for diagnosing small bowel diseases were evaluated using enteroscopy results as the standard. RESULTS: Among the 576 children, small bowel lesions were detected by enteroscopy in 274 children (47.6%).There was no significant difference in lesion detection rates or complication rates between the SBE and DBE groups (P>0.05), but the DBE group had deeper insertion, longer procedure time, and higher complete small bowel examination rate (P<0.05). The complication rate during enteroscopy was 4.3% (25/576), with 18 cases (3.1%) of mild complications and 7 cases (1.2%) of severe complications, which improved with symptomatic treatment, surgical, or endoscopic intervention. Among the 412 children who underwent CTE, the sensitivity and specificity for diagnosing small bowel diseases were 44.4% and 71.3%, respectively. CONCLUSIONS SBE and DBE have similar diagnostic efficacy for pediatric small bowel diseases, but DBE is preferred for suspected deep small bowel lesions and comprehensive small bowel examination. Enteroscopy in children demonstrates relatively good overall safety. CTE demonstrates relatively low sensitivity but comparatively high specificity for diagnosing small bowel diseases.
Retrospective Studies ; Treatment Outcome ; Double-Balloon Enteroscopy/statistics & numerical data* ; Single-Balloon Enteroscopy/statistics & numerical data* ; Humans ; Male ; Female ; Child ; Operative Time ; Tomography, X-Ray Computed/statistics & numerical data* ; Sensitivity and Specificity ; Intestine, Small/surgery* ; Intestinal Diseases/surgery*

OBJECTIVES: To study the effect of prophylactic phenytoin (PHT) or levetiracetam (LEV) on busulfan (BU) blood concentration in children undergoing hematopoietic stem cell transplantation. METHODS: Pediatric patients conditioned with BU plus cyclophosphamide and fludarabine at the First People's Hospital of Chenzhou from September 2023 to February 2025 were retrospectively included. Patients were grouped by prophylactic antiepileptic regimen into PHT (n=24) and LEV (n=26). BU blood concentrations at the end of infusion (0 hour) and at 1, 2, and 4 hours post-infusion were compared between groups. RESULTS: At 0 hour post-infusion, BU blood concentrations did not differ significantly between groups (P>0.05). At 1, 2, and 4 hours post-infusion, BU blood concentrations were higher in the LEV group than in the PHT group (P<0.05). The area under the concentration-time curve from 0 to ∞ (AUC_0-∞) was greater in the LEV group (P<0.001), and the attainment rate of AUC_0-∞ was higher in the LEV group than in the PHT group (73% vs 21%, P<0.001). No significant differences were observed between groups in time to hematopoietic engraftment or in the incidence of BU-related adverse drug reactions (P>0.05). CONCLUSIONS Compared with PHT, LEV prophylaxis is associated with higher BU blood concentration and a higher AUC_0-∞ attainment rate. There is no observed difference in BU efficacy or safety between PHT and LEV.
Humans ; Levetiracetam/therapeutic use* ; Busulfan/pharmacokinetics* ; Hematopoietic Stem Cell Transplantation ; Male ; Female ; Child ; Child, Preschool ; Phenytoin/pharmacology* ; Infant ; Retrospective Studies ; Anticonvulsants/pharmacology* ; Adolescent

ObjectiveTo analyze the effects of new integration processing method in producing area and traditional method on the composition and pharmacological action of Polygoni Multiflori Radix Praeparata（PMRP）， and to illustrate the advantages of toxicity reducing and efficacy enhancing of the decoction pieces prepared by the new method. MethodFresh Polygoni Multiflori Radix（PMR） was taken from Dao-di producing area， and was processed by new integration processing method in producing area（steaming with black bean juice under pressure of 0.1 MPa and temperature at 120 ℃ for 10.5 h） and traditional method（steaming with black bean juice under water for 36 h）， respectively. Samples were collected during the processing process of the two methods， For new method， the samples were collected at 0.5， 3， 5.5， 8， 10.5 h， separately. For traditional method， the samples were collected every 4 h. High performance liquid chromatography（HPLC） was used to establish fingerprint and identify common peaks， the content of polysaccharides was determined by anthrone-sulfuric acid colorimetry at 627 nm， and the contents of anthraquinones and stilbene glycosides in different processed products were determined according to the methods under the item of determination of PMR and PMRP in the 2020 edition of Chinese Pharmacopoeia. In pharmacological experiments， 90 SD rats were randomly divided into 9 groups with 10 in each group（half of male and half of female）， including the blank group， and raw products， 24 h processed products under atmospheric pressure， 30 h processed products under atmospheric pressure， 8 h processed products under high pressure groups with low and high dosages（4.125， 16.5 g·kg^-1）. Rats were given the drug by gavage for 29 d with once a day， blood was collected from the abdominal aorta after the last administration， and the serum was isolated， the body mass and liver mass of rats were weighed and the organ index was calculated. The pathological change of liver tissue was observed by hematoxylin-eosin（HE） staining， and biochemical methods were used to detect the contents of aspartate aminotransferase（AST）， alanine aminotransferase（ALT）， alkaline phosphatase（ALP）， γ-glutamyltransferase（GGT）， lactic dehydrogenase（LDH） in serum which used as liver function indicators and the levels of superoxide dismutase（SOD）， malondialdehyde（MDA）， glutathione peroxidase（GSH-Px） in brain tissues which used as oxidation indicators. ResultA total of 14 common peaks were identified in the fingerprint of PMR， PMRP prepared by new method and traditional method， and three of the peaks were designated as stilbene glycoside， emodin and emodin methyl ether， respectively. The characteristic peak areas of each processed products changed significantly from 0 min to 25 min， indicating that different processing methods had an effect on the contents of components with high polarity in PMRP， and the trend of the changes of the two methods was similar， with the higher degree of change in the new method. The determination results showed that compared with the traditional method， the content of polysaccharide（a kind of beneficial component in PMRP obtained by the new method） significantly increased， while the contents of stilbene glycoside and bound anthraquinone（liver-damaging ingredients） significantly decreased. The pharmacological results showed that compared with the blank group， AST and LDH levels of male rats in the low and high dose groups of 24 h processed products under atmospheric pressure and AST level of male rats in the low and high dose groups of 8 h processed products under high pressure were significantly reduced（P<0.05， P<0.01）， while compared with the raw product groups with the same dose， AST and LDH levels of male rats in the low dose group of 30 h processed products under atmospheric pressure were significantly reduced（P<0.05， P<0.01）， the AST levels of male rats in the low and high dose groups of 8 h processed products under high pressure were significantly decreased（P<0.01）， and there was no statistical significance in the differences of biochemical indexes of female rats in each administration group as compared with those of the blank group. ConclusionThe new integration processing method in producing area of PMRP can reach the quality of relevant regulations in 8 h. The processed products obtained by this method have more advantages than the traditional method in terms of toxicity reducing and efficacy enhancing， and energy saving to avoid the loss of ingredients， which can provide ideas for the production of high-quality decoction pieces of PMRP， and the integration processing method in producing area of other roots and rhizomes of traditional Chinese medicines.

BACKGROUND:Inflammation is one of the important factors that induce cerebral ischemia-reperfusion injury.Studies have shown that electroacupuncture can effectively reduce inflammation after ischemic stroke and improve the symptoms of neurological deficits,but the mechanism is not clear. OBJECTIVE:To observe the effect of electroacupuncture on Toll-like receptor 4/nuclear factor-κB in rats with cerebral ischemia-reperfusion injury. METHODS:Forty-eight male Sprague-Dawley rats were randomly divided into sham operation group,model group and electroacupuncture group,with 16 rats in each group.The rat model of cerebral ischemia-reperfusion injury was prepared by middle cerebral artery occlusion.At 24 hours after modeling,the rats in the electroacupuncture group were treated with electroacupuncture,once a day,20 minutes each time,for a total of 5 days.The sham operation group and the model group did not do any intervention.After 5 days of intervention,Longa method was used to evaluate the degree of neurological injury in rats.Triphenyl tetrazolium chloride staining and hematoxylin-eosin staining were used to measure the volume of cerebral infarction and the pathological changes of brain tissue in rats.Serum interleukin-6,interleukin-18 and tumor necrosis factor-α were detected by ELISA.Expressions of Toll-like receptor 4 and nuclear factor-κB in the cerebral cortex at mRNA and protein levels were detected by fluorescence quantitative PCR and western blot,respectively. RESULTS AND CONCLUSION:Compared with the sham operation group,the neurological function scores,serum interleukin-6,interleukin-18,and tumor necrosis factor-α levels,Toll-like receptor 4 and nuclear factor-κB mRNA and protein expression levels were significantly higher in the model group(P＜0.01).Compared with the model group,electroacupuncture significantly reduced the neurological function scores,serum interleukin-6,interleukin-18,and tumor necrosis factor-α levels,Toll-like receptor 4 and nuclear factor-κB mRNA and protein expression levels(P＜0.05,P＜0.01).Compared with the sham operation group,the volume of cerebral infarction in the model group increased significantly(P＜0.01).Compared with the model group,the volume of cerebral infarction in the electroacupuncture group decreased(P＜0.05).In the model group,the arrangement of neurons was disordered,some nerve cells disappeared,nuclei presented with pyknosis and incomplete structure.After electroacupuncture intervention,the degree of neuronal degeneration and neuronal loss in the cerebral cortex of rats were reduced compared with those in the model group.To conclude,electroacupuncture can significantly improve the neurobehavior of rats with cerebral ischemia-reperfusion injury,reduce brain tissue injury,and effectively reduce the level of serum inflammatory factors.The mechanism may be related to the inhibition of Toll-like receptor 4/nuclear factor-κB signaling pathway.

Humans ; Consanguinity ; Exome Sequencing ; Mutation/genetics* ; Sequence Analysis, DNA ; Ciliary Motility Disorders ; Axonemal Dyneins/genetics*

AIM To investigate the variation rules of main secondary metabolites in Hedysari Radix before and after rubbing strip.METHODS UPLC-MS/MS was adopted in the content determination of formononetin,ononin,calycosin,calycosin-7-glucoside,medicarpin,genistein,luteolin,liquiritigenin,isoliquiritigenin,vanillic acid,ferulic acid,γ-aminobutyric acid,adenosine and betaine,after which cluster analysis,principal component analysis and orthogonal partial least squares discriminant analysis were used for chemical pattern recognition to explore differential components.RESULTS After rubbing strip,formononetin,calycosin,liquiritigenin and γ-aminobutynic acid demonstrated increased contents,along with decreased contents of ononin,calycosin-7-glucoside and vanillic acid.The samples with and without rubbing strip were clustered into two types,calycosin-7-glucoside,formononetin,γ-aminobutynic acid,vanillic acid,calycosin-7-glucoside and formononetin were differential components.CONCLUSION This experiment clarifies the differences of chemical constituents in Hedysari Radix before and after rubbing strip,which can provide a reference for the research on rubbing strip mechanism of other medicinal materials.