AIM To establish an HPLC method for the simultaneous content determination of 5-hydroxymethylfurfural,chlorogenic acid,caffeic acid,strychnine,paeoniflorin,ferulic acid,paeoniflorin Ⅰ,epimedium glycoside,psoralen,isopsoralen and glycyrrhetinic acid in Bunao Soft Capsules.METHODS The analysis was performed on a 35 ℃ thermostatic Waters Symmetry C18 column(250 mm×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 230,280 nm.RESULTS Eleven constituents showed good linear relationships within their own ranges(r＞0.999 0),whose average recoveries were 98.47％-103.30％with RSDs of 1.13％-2.80％.CONCLUSION This simple and reliable method can be used for the quality control of Bunao Soft Capsules.

Objective:To investigate the role of SiHa cell-derived exosomal hsa-miR-29c-3p in the angiogenesis of cervical cancer(CC).Methods:Cancer tissue specimens from 45 patients with cervical squamous cell carcinoma(CSCC)and normal cervical tissue specimens from 15 controls were collected from Department of Gynecology,Hengyang Central Hospital from January 2019 to December 2021.CSCC SiHa cells and human umbilical vein endothelial cells(HUVECs)were routinely cultured.miRNA-NC,hsa-miR-29c-3p,si-miRNA-NC,and si-hsa-miR-29c-3p were transfected into SiHa cells with Lipofectamine 2000,grouped as miRNA-NC group,hsa-miR-29c-3p group,si-miRNA-NC group and si-hsa-miR-29c-3p group,respectively.HUVECs were transfected with mimic-NC,miR-29c-3p-mimic,pCMV-NC,and pCMV-ATAD2B(ATPase family protein 2B with AAA domain)using Lipofectamine 2000,grouped as the mimic-NC group,miR-29c-3p-mimic group,pCMV-NC group,pCMV-ATAD2B group,and pCMV-ATAD2B+miR-29c-3p-mimic group.The expression of hsa-miR-29c-3p in CSCC tissues was detected by in situ hybridization(ISH),and CD31-positive blood vessels in CSCC tissues and xenograft tissues were detected by immunohistochemistry(IHC).Exosomes from SiHa and C33a cells were isolated and characterized using transmission electron microscopy(TEM)and western blotting(WB).The uptake of exosomes by HUVECs was examined.The expression of hsa-miR-29c-3p and ATAD2B mRNA in SiHa and C33a cells,as well as in their derived exosomes,was detected using qPCR.Tube-forming assay,Transwell assay,and scratch healing assay were performed to detect the effect of exosomes on the ability of HUVEC migration and tube formation.Dual luciferase reporter gene assay verified the interaction between hsa-miR-29c-3p and ATAD2B.Xenograft experiments examined the effects of SiHa cell-derived exosomes on transplanted tumor growth and angiogenesis in each group.Results:hsa-miR-29c-3p was highly expressed in CSCC tissues and was positively correlated with microvessel density(MVD)(all P<0.05).Exosomes derived from SiHa and C33a cells exhibited typical exosomal morphology and protein expression patterns.Exosomal hsa-miR-29c-3p from SiHa and C33a cells were efficiently taken up by HUVECs in vitro.The SiHa cell-derived exosomal hsa-miR-29c-3p promoted not only the tube-forming and migration of HUVECs in vitro but also the xenograft growth and angiogenesis in vivo(all P<0.05).hsa-miR-29c-3p directly targeted ATAD2B and regulated its expression(P<0.05).Overexpression of ATAD2B reversed the promotive effect of hsa-miR-29c-3p on tube-formation,migration,and scratch-healing in HUVECs(all P<0.05).Conclusion:SiHa cell-derived exosomal hsa-miR-29c-3p regulates angiogenesis in CSCC tissues by targeting ATAD2B.Exosomal hsa-miR-29c-3p may be a potential diagnostic marker and therapeutic target for CC diagnosis and treatment.

AIM To study the chemical constituents from Pyrolae Herba and their activity against pathogenic bacteria associated with sports injuries.METHODS Silica gel and Sephadex LH-20 were used for isolation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The antibacterial activity was detected by filter paper method.RESULTS Twenty five compounds were isolated and identified as manuleoside H(1),5-hydroxy-3,7-dimethoxy-4,-methyl flavone(2),5-hydroxy-3,7,4'-methoxy-flavone(3),5-hydroxy-7,3',4'-trimethoxy-flavone(4),5,7-dihydroxy-3-methoxy-flavone(5),7.5,4'-trihydroxyflavone(6),ethyl vanillate(7),clovandiol(8),hydroxydihydrobovolide(9),zanthopyranone(10),eugenin(11),berchemol(12),3,7-dimethoxy-5,3',4'-trihydroxyflavone(13),2,5-dimethyl-7-hydroxychromone(14),rutalinium(15),formononetin(16),8-hydroxy-4',7-dimethoxy-isoflavones(17),isoline(18),dearabinosyl pneumonanthoside(19),3,5-dihydroxycinnamic acid(20),2,4,6-trihydroxyacetophenone(21),p-isopropyl benzoic acid(22),geraldone(23),ethylsyringin(24),4-hydroxyphenylethyl-2'-hydroxypropionate(25).Compounds 8,10,12,16 and 24 showed bacteriostatic zone on pathogenic bacteria susceptible to sports injuries.CONCLUSION All the compounds are isolated from Pyrolae Herba for the first time.Compounds 8,10,12,16 and 24 have inhibitory activity against pathogenic bacteria susceptible to sports injuries.

AIM To investigate the clinical effects of Huoxue Qingre Formula combined with radiofrequency ablation on patients with thyroid nodules due to Phlegm Stagnation and Blood Stasis.METHODS Ninety patients were randomly assigned into control group(45 cases)for 3-month intervention of both Huoxue Qingre Formula placebo and radiofrequency ablation,and observation group(45 cases)for 3-month intervention of both Huoxue Qingre Formula and radiofrequency ablation.The changes in clinical effects,VRR,TCM syndrome score,cytokines(TNF-α,IFN-γ,IL-23,IL-1β),biochemical indices[Ca2+,25(OH)D,CRP],thyroid function indices(TSH,FT3,FT4)and safety indices were detected.RESULTS At postoperative 12 months,the observation group demonstrated higher total effective rate than the control group(P＜0.05).At postoperative 3-12 months,the two groups displayed increased VRR(P＜0.05),especially for the observation group(P＜0.05,P＜0.01).At the 7th,30th postoperative days,the two groups displayed decreased TCM syndrome score and cytokines(P＜0.05,P＜0.01),especially for the observation group(P＜0.05,P＜0.01).At the 30th postoperative day,the two groups displayed decreased Ca2+(P＜0.01)and increased 25(OH)D(P＜0.01),especially for the observation group(P＜0.01).At the 7th postoperative day,the two groups displayed increased CRP(P＜0.01),especially for the control group(P＜0.05),which were decreased at the 30th postoperative day(P＜0.05,P＜0.01),especially for the observation group(P＜0.01).No significant differences in thyroid function indices and safety indices were found between the two groups(P＞0.05).CONCLUSION For the patients with thyroid nodules due to Phlegm Stagnation and Blood Stasis,Huoxue Qingre Formula combined with radiofrequency ablation can safely and effectively enhance nodule VRR,alleviate discomfort symptoms,reduce TCM syndrome score,inflammatory factor levels,and accelerate recovery.

Objective To investigate the distribution patterns of TCM syndrome elements and syndrome types in Klebsiella pneumoniae pneumonia.Methods The study adopted a retrospective research method to collect the basic demographic data,pathogenic characteristics,TCM syndrome patterns,and four diagnostic examination information(including tongue and pulse manifestations)of hospitalized patients with Klebsiella pneumoniae pneumonia at Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,from September 2022 to June 2024.Based on clustering analysis and factor analysis,the study explored the characteristics of TCM syndrome elements and the distribution patterns of TCM syndromes in Klebsiella pneumoniae pneumonia.Results A total of 365 patients(236 males,129 females)aged 23-100 years were included.Statistical analysis identified 33 core syndrome items,with cough being the most frequent(66.0%),followed by viscous phlegm(42.2%);the most common tongue characteristics were red tongue(50.4%)and greasy coating(45.8%),with slippery pulse being predominant(54.0%).Clustering analysis revealed four syndrome types:lung-kidney yin deficiency,phlegm-dampness with qi deficiency,yang deficiency with water overflow,and phlegm-heat obstructing the lung.Phlegm-heat obstructing the lung was the most prevalent(38.1%),followed by yang deficiency with water overflow(22.5%)and lung-kidney yin deficiency(22.2%).Factor analysis identified four common factors encompassing 21 variables,with a cumulative variance contribution rate of 74.888%.The results of syndrome elements were divided into phlegm,heat,qi stagnation,water retention,dampness,yang deficiency,yin deficiency,qi deficiency,and lung,kidney,heart,spleen based on pathological syndrome elements and pathological location syndrome elements.Conclusion Klebsiella pneumoniae pneumonia primarily affects the lungs,kidneys,heart,and spleen,with the lungs being the key organ.The most common TCM syndrome types are phlegm-heat obstructing the lung,lung-kidney yin deficiency,yang deficiency with water overflow,and phlegm-dampness with qi deficiency,among which phlegm-heat obstructing the lung is the most prevalent.

Objective:To investigate the regulatory effects of transcutaneous auricular vagus nerve stimulation (taVNS) on functional connectivity (FC) of thalamic subregions in patients with premenstrual syndrome (PMS).Methods:This study was a cross-sectional investigation. Clinical, laboratory, and imaging data were retrospectively collected from 56 PMS patients (PMS group) and 66 healthy controls (control group) recruited from various universities and hospitals in Nanning between November 2021 and June 2024. Resting-state functional MRI (fMRI) data and fMRI data during taVNS immediate stimulation (2 Hz, 25 Hz) were acquired from subjects during their late luteal phase. Using thalamic subregions (anterior thalamic nucleus, lateral nucleus, ventral nucleus, medial nucleus, central nucleus, posterior nucleus) as seeds, two-sample t-tests or paired t-tests were employed to analyze alterations in thalamic subregion FC in PMS patients and the regulatory effects of taVNS on these changes. Independent samples t-test were used to compare the differences in clinical and laboratory indicators between the PMS group and the control group. The relationship between taVNS regulation of thalamic subregion FC in PMS patients and thalamic internal functional connectivity were analyzed using mediation effect analysis. Results:Compared to the control group, patients in the PMS group showed increased scores on the Daily Record of Severity of Problems, Pittsburgh Sleep Quality Index, Self-Rating Anxiety Scale, Self-Rating Depression Scale, Hamilton Anxiety Rating Scale 17, and Hamilton Depression Rating Scale 14 during the late luteal phase ( P<0.05). At baseline, PMS patients exhibited higher FC between the left thalamic lateral nucleus and the left insula, and lower FC between the left medial nucleus, posterior nucleus, and ventral nucleus of the thalamus and the right middle frontal gyrus (MFG) compared to the control group (GRF corrected, voxel-level P<0.001, cluster-level P<0.05). During 2 Hz taVNS immediate stimulation in PMS group, FC between the left thalamic medial nucleus, posterior nucleus, ventral nucleus and the right MFG, as well as the FC between the left thalamic ventral nucleu and the left MFG increased compared to baseline levels; meanwhile, FC between the left thalamic posterior nucleus, ventral nucleus and the left insula decreased compared to baseline levels (GRF corrected, voxel-level P<0.001, cluster-level P<0.05). During 25 Hz taVNS immediate stimulation, the FC between the left thalamic ventral nucleus and the right MFG decreased compared to the baseline level (GRF corrected, voxel-level P<0.001, cluster-level P<0.05). Mediation effect analysis showed that the FC between the left thalamic posterior nucleus and the left lateral nucleus mediated part of the association between the FC of the left lateral thalamic nucleus-left insula and the FC of the left ventral thalamic nucleus-left putamen/insula; there were significant direct effects between the FC of the left lateral thalamic nucleus-the left posterior nucleus and FC of the left lateral thalamic nucleus-the left insula, as well as between the FC of the left ventral thalamic nucleus-the left MFG and FC of the left ventral thalamic nucleus-the right MFG. Conclusions:taVNS can modulate abnormal FC of the left thalamic subregions in PMS patients, restoring it toward normalization. The regulatory effects of 2 Hz stimulation are more pronounced than those of 25 Hz stimulation. This modulation primarily operates through two pathways: the left thalamic lateral nucleus-left insula-left thalamic ventral nucleus pathway and the left MFG-left thalamic ventral nucleus-right MFG.

A centrifugation-free,single-reaction colorimetric method for detection of glucose,utilizing a dual nanozyme cascade system based on gold nanoparticles(AuNPs)and iron-doped carbon dots(FeCDs),was developed in this work.The AuNPs exhibited glucose oxidase-like activity to catalyze glucose oxidation for generation of H2O2,while the FeCDs demonstrated peroxidase-like activity to subsequently catalyze the H2O2-mediated oxidation of 3,3',5,5'-tetramethylbenzidine(TMB).To prevent interference from the blue signal generated by self-aggregation of AuNPs in subsequent quantitative detection,the reaction system was terminated with HCl,converting oxTMB into a stable yellow product.Based on changes in the absorbance at 450 nm of this yellow solution,a quantitative relationship was established between glucose concentration and absorbance at 450 nm(A450).Experimental results demonstrated that this sensor achieved a linear detection range of 44 μmol/L to 11.11 mmol/L(R2=0.993)with a detection limit of 30.68 μmol/L and spiked recoveries of 97.9%-104.7%.By integrating smartphone-based color recognition capabilities,a rapid visual detection platform was established for quantification of glucose through RGB analysis.The validation experimental results using commercial glucose injection samples further confirmed the practical application potential of this methodology.

Water-soluble silver nanoclusters(DHLA-AgNCs)with red fluorescence emission were synthesized using silver nitrate(AgNO3)as silver source,dihydrolipoic acid(DHLA)as ligand and sodium borohydride(NaBH4)as reducing agent by one-pot method.Based on the selective quenching of DHLA-AgNCs by tetracycline(TC),a rapid and selective fluorescence analysis method for detection of TC was constructed by monitoring the fluorescence intensity change at 650 nm.Under the optimal detection conditions,the fluorescence quenching efficiency of DHLA-AgNCs showed good linear relationship with concentration of TC within the range of 10.0-120.0 μmol/L,and the limit of detection(LOD)was 0.39 μmol/L.This method was successfully applied to detection of TC in milk samples,with spiked recoveries ranging from 99.5%to 102.5%,and relative standard deviations(RSDs)less than 5%.This method had the advantages of simplicity,rapidity,strong specificity and low cost,and thus provided a simple and feasible strategy for selective detection of TC.

Nano/microplastics(NMPs),due to their environmental persistence and resistance to degradation,have emerged as a major contributor to global pollution.NMPs are capable of adsorbing various hazardous chemicals and heavy metals,thereby posing threats to aquatic ecosystem health,which may ultimately cause potential risks to human health.Conventional analytical methods suffered from limited resolution,insufficient chemical information,or destruction of sample,invalidating these assays for on-site detection of NMPs.Surface-enhanced Raman scattering(SERS)offers distinct advantages such as high sensitivity,superior specificity,rich fingerprint information,and non-destructive analysis,thus facilitating the on-site analysis of NMPs in complex matrices.This review summarized recent advances in SERS substrates for detection of NMPs,discussed the construction and applications of SERS-based multimodal detection strategies,and introduced the research progress of SERS detection of NMPs in food safety,environmental pollution,and bioanalysis.Moreover,the main challenges and future directions of SERS-based NMP detection were outlined.

A portable molecularly imprinted(MIP)surface-enhanced Raman scattering(SERS)chip was fabricated via a green electrochemical approach for highly selective detection of bisphenol A(BPA).This MIP-AuNP/UIO-66/SPE sensor was fabricated through a single-step co-deposition process.The process involved electropolymerization onto a UIO-66 modified screen-printed electrode(SPE),by usingo-phenylenediamine(OPD)as functional monomer and BPA as template,and simultaneously electro-reduction generated gold nanoparticles(AuNPs),which served as the SERS-active substrate.Ultimately,this one-step method formed a three-dimensional porous architecture on the electrode surface.Under 785 nm laser excitation,the sensing chip exhibited a highly sensitive SERS response towards BPA.The intensity of its characteristic peak at 850 cm-1 showed a good linear relationship with logarithm of BPA concentration in the range of 1.0×10-10 to 1.0×10-6 mol/L,with a detection limit of 1.0×10-12 mol/L.More importantly,the fabricated chips maintained highly selective binding affinity for BPA in water samples even in the presence of structural analogs bisphenol F(BPF)and bisphenol S(BPS).When the chip was applied to detection of BPA in water samples from plastic bottle and paper cup,the recovery rates ranged from 94.0%to 103.0%with relative standard deviations(RSD)less than 4.7%.The developed chip offered a highly sensitive and selective solution for detection of trace BPA in complex water samples.