This study aims to explore the scientific connotation of sinking Rehmanniae Radix has the best quality and compare the quality between floating and sinking fresh Rehmanniae Radix samples. Ultra-performance liquid chromatography tandem quadrupole electrostatic field Orbitrap high-resolution mass spectrometry(UHPLC-Q-Orbitrap HRMS) was employed to detect the chemical components in floating and sinking fresh Rehmanniae Radix samples. The fingerprint of fresh Rehmanniae Radix was established by high performance liquid chromatography(HPLC), and four index components were determined simultaneously. The cluster analysis, principal component analysis(PCA), and orthogonal partial least squares-discriminant analysis(OPLS-DA) were conducted to compare the quality of floating and sinking fresh Rehmanniae Radix samples. An evaporative light-scattering detector was used to compare the content of five sugars. The extract yield and drying rate were determined, and the quality connotation of sinking Rehmanniae Radix has the best quality was explained by multiple indicators. A total of 41 components were preliminarily identified from fresh Rehmanniae Radix by UHPLC-Q-Orbitrap HRMS, including 7 iridoid glycosides, 9 phenylethanol glycosides, 6 amino acids, 4 sugars, 3 phenolic acids, 5 nucleosides, 3 organic acids, 1 ionone, 1 furan, 1 coumarin, and 1 phenylpropanoid. The results showed that the main chemical components were consistent between floating and sinking fresh Rehmanniae Radix. Nine common peaks were identified in the fingerprints of 15 batches of floating and sinking fresh Rehmanniae Radix samples, and the similarity of fingerprints was greater than 0.9. The cluster analysis, PCA, and OPLS-DA classified floating and sinking fresh Rehmanniae Radix sasmples into two categories, indicating differences in the quality between them. The total content of catalpol, rehmannioside D, ajugol, and verbascoside in sinking fresh Rehmanniae Radix samples was higher than that in floating samples of the same batch and specification, and the main differential component was catalpol. The total content of fructose, glucose, sucrose, raffinose, and stachyose in sinking fresh Rehmanniae Radix samples was higher than that in floating samples of the same batch and specification, and the main differential component was stachyose. The extract yield and drying rate of the sinking samples were higher than those of floating samples. This study preliminarily showed that floating and sinking fresh Rehmanniae Radix samples had the same components but great differences in the content of medicinal substance basis. The total content of four glycosides and five sugars, extract yield, and drying rate of sinking fresh Rehmanniae Radix samples is higher than that of floating samples of the same batch and specification. These findings, to a certain extent, explains the scientificity of sinking Rehmanniae Radix has the best quality recorded in ancient books and provide a reference for the quality control and clinical application of fresh Rehmanniae Radix.
Chromatography, High Pressure Liquid/methods* ; Drugs, Chinese Herbal/chemistry* ; Rehmannia/chemistry* ; Chemometrics ; Mass Spectrometry/methods* ; Quality Control ; Principal Component Analysis ; Plant Extracts

Data analysis models may assist the transmission of traditional Chinese medicine （TCM） experience and clinical diagnosis and treatment， and the possibility of constructing a “data-knowledge” dual-drive model was explored by taking gastric precancerous state as an example. Data-driven is to make clinical decisions around data analysis， and its syndrome-differentiation decision-making research relies on hidden structural models and partially observable Markov decision-making processes to identify the etiology of diseases， syndrome elements， evolution of pathogenesis， and syndrome differentiation protocols； knowledge-driven is to make use of data and information to promote decision-making and action processes， and its syndrome-differentiation decision-making research relies on convolutional neural networks to improve the accuracy of local disease identification and syndrome differentiation. The “data-knowledge” dual-driven model can make up for the shortcomings of single-drive numerical simulation accuracy， and achieve a balance between local disease identification and macroscopic syndrome differentiation. On the basis of previous research， we explored the construction method of diagnostic assisted decision-making platform for gastric precancerous state， and believed that the diagnostic and decision-making ability of doctors can be extended through the assistance of machines and algorithms. Meanwhile， the related research methods were integrated and the core features of gastric precancerous state based on TCM syndrome differentiation and endoscopic pathology diagnosis and prediction were obtained， and the elements of endoscopic pathology recognition based on TCM syndrome differentiation were explored， so as to provide ideas for the in-depth research and innovative application of cutting-edge data analysis technology in the field of intelligent TCM syndrome differentiation.

Objective To investigate the role of variation trend of pre-treatment blood beta human chorionic gonadotropin(β-HCG)in predicting the efficacy of tubal pregnancy(TP)treated by drug therapy.Methods Clinical data of 380 TP patients treated by drug therapy at the First Affiliated Hospital of Guangzhou University of Chinese Medicine from January 2017 to June 2022 were collected.The patients were divided into 271 cases in the pure Chinese medicine(CM)treatment group(shortened as pure CM group)and 109 cases in the combination of Chinese medicine and western medicine group(shortened as combination group)according to the therapy.The relevant factors that may affect the outcomes of the therapy were analyzed,and logistic regression analysis was used for the control of confounding factors.The influences of pre-treatment β-HCG on the outcomes of TP treated by various therapies were investigated,and the value of variation trend of pre-treatment β-HCG in predicting the efficacy of TP treated by drug therapy was assessed using the receiver operating characteristic curve(ROC curve).Results(1)The results of univariate analysis showed that the initial value of blood β-HCG,the variation trend of β-HCG and the difference value of β-HCG detected for 2 times before treatment were correlated with the treatment outcomes in the pure CM group and the combination group,and the differences were all statistically significant(P＜0.05 or P＜0.01).(2)The results of multivariate regression analysis showed that in the pure CM group,the blood β-HCG values and the variation trend of β-HCG detected for 2 times before treatment were correlated with the treatment outcomes,and the differences were all statistically significant(P＜0.05 or P＜0.01);in the combination group,the menopausal time and blood β-HCG value detected at the second time before treatment were correlated with the treatment outcomes,and the differences were all statistically significant(P＜0.05).(3)After controlling for the confounding factors,the variation trend of pre-treatment β-HCG in the pure CM group was correlated with the treatment outcomes,and the difference was statistically significant(OR=3.35,95%CI being 1.69-6.65,P = 0.001).However,the variation trend of pre-treatment β-HCG in the combination group did not affect the treatment outcomes,and the difference was not statistically significant(P＞0.05).(4)The ROC curve analysis showed that the area under the curve(AUC)in the pure CM group was 0.74,the cutoff value was-0.14,the sensitivity was 71.3%and the specificity was 67.8%.The AUC in the combination group was 0.67,the cutoff value was 0.10,the sensitivity was 73.6%and the specificity was 60.7%.Conclusion The variation trend of pre-treatment blood β-HCG can effectively predict the efficacy of Chinese medicine for the treatment of TP,but has no value for predicting the efficacy of the combination of Chinese medicine and western medicine in treating TP.The variation value of pre-treatment blood β-HCG can be used as a valuable reference indicator for the early prediction of the outcomes of TP treated by drug therapy.

Hydrogen sulfide(H2S)is an emerging endogenous neuromodulator that holds significant potential in the realm of neurological diseases.Its role encompasses reducing neuronal damage,inhibiting excessive activation of striatal astrocytes,and regulating cerebrovascular function,among other physiopathological pathways.Conversely,microRNAs(miRNAs)are widely recognized as pivotal regulators of neurological diseases.They primarily target the 3'untranslated region of the target gene mRNA,impeding mRNA translation and hindering its expression to impart neuroprotective effects.Recent findings have underscored the crucial involvement of H2S and miRNAs in the pathogenesis of various neurological disorders such as Parkinson's disease,stroke,and spinal cord diseases,thereby garnering significant attention.This paper comprehensively summarizes the protective effects arising from the interplay between H2S and miRNAs in neurological diseases,while also delving into the potential therapeutic efficacy they hold for such conditions.

Objective:To prepare a humanized monoclonal antibody against periostin and establish a stable cell line.Meth-ods:Based on anti-periostin mouse monoclonal antibody developed by our laboratory,total RNA was extracted,and variable region sequences were obtained by RT-PCR amplification of VH and VL genes.The mouse antibody CDR region was transplanted into the human antibody framework receptor region,and the gene was subcloned into the expression vector PATX-GS2,and stably transfected into CHO cells.Monoclonal cell lines were obtained by MSX pressure screening and limited dilution.Results:VH and VL genes were amplified by RT-PCR,and the sequence of the light and heavy chain variable region were determined.Antibody humanization were successfully stablished by CDR transplantation method a murine antibody to a human framework,and a eukaryotic expression plasmid was constructed,which was transfected into CHO cells for expression,and human anti-periostin antibody was successfully obtained.ELISA and Western blot results showed that the humanized antibody had good anti-periostin activities and binding affinity.Conclu-sion:In this study,anti-periostin humanized monoclonal antibody has been successfully prepared,which can specifically bind to peri-ostin proteins in vivo and have biological activity,providing scientific data for the precise treatment of retinal fibrosis,tissue and organ fibrosis,and malignant tumors.

Objective To compare the effects of proprioceptive neuromuscular facilitation(PNF),con-centric-isometric-eccentric fast training(CIEFT),and concentric-isometric-eccentric slow training(CI-EST)in young adults with flexible flatfoot.Methods Forty participants were randomly allocated into a PNF group,a CIEFT group,a CIEST group and a control group,each of 10.The PNF,CIEFT and CIEST groups underwent six weeks of training as their group names indicated.Then the height differ-ence in the navicular drop test(NDt),normalized navicular height truncated(NNHt),muscle strength of ankle joint,plantar pressure distribution characteristics,and dynamic balance of all groups were re-corded before and after the intervention.Results As to the arch morphology,compared with pre-inter-vention,NDt decreased significantly in the dominant side of PNF group(P=0.049)and the non-domi-nant side of CIEFT group(P=0.034),while NNHt increased significantly in the non-dominant side of CIEFT group(P=0.026).Moreover,compared with pre-intervention,the muscle strength of plantar flex-ion increased significantly in all groups except the control group(dominant/non-dominant side:P=0.003/P=0.004,P=0.000/P=0.000,P=0.001/P=0.001),and that of inversion increased significantly in both PNF and CIEFT groups(dominant/non-dominant side:P=0.011/P=0.005,P=0.003/P=0.003).When it comes to the plantar pressure distribution characteristics,after the intervention,in the non-dominant side,the incremental center of pressure(COP)connections decreased significantly in CIEFT group(P=0.037),while the ratio of medial arch load and the contact area of the medial arch in PNF group de-creased significantly(P=0.012,P=0.027).Moreover,in the dominant side,the contact area decreased significantly in CIEST group(P=0.038),but increased significantly in the control group(P=0.015).What's more,after intervention,the Y-balance test score increased significantly in both sides of PNF and CIEST groups and the dominant side of CIEFT group(P=0.006/P=0.023,P=0.001/P=0.035,P=0.011).Conclusion Through a 6-week exercise intervention,PNF can improve the foot arch morphology and enhance dynamic balance ability in young adults with flexible flatfoot,while concentric-isometric-eccen-tric fast and slow training is superior in improving the foot arch morphology and the dynamic balance ability,respectively.

Aim To investigate the effect and role of neuronal restriction silencing factor(REST/NRSF)in epilepsy disorder.Methods Immunohistochemistry,immunofluorescence,Western blot and qPCR tech-niques were used to detect REST/NRSF expression levels in hippocampal tissues of mice induced by kainic acid and human brain tissue.Viral injections,EEG re-cordings and behavioral methods were used to test the effects on epileptic mice after knockdown and overex-pression of REST/NRSF in the hippocampal CA1 re-gion,respectively.Results The positive rate of REST/NRSF in the lesions of epileptic patients was significantly higher compared with that in the control group.The levels of REST/NRSF protein and mRNA in the CA1 region of the hippocampus of mice in the KA model group were significantly higher.Kv7.2 and Kv7.3 potassium channel mRNA expression levels were significantly down-regulated.Significant up-regu-lation of REST/NRSF expression levels was observed in mouse hippocampus after NMDA injection.Knock-down of REST/NRSF in the CA1 region of hippocam-pus significantly elevated the expression levels of Kv7.2 and Kv7.3 potassium channel mRNAs.The fre-quency of EEG spiking and sharp-wave issuance and epileptic seizure grade were significantly lower.Over-expression of REST/NRSF in the CA1 region of hippo-campus significantly reduced the mRNA expression lev-els of Kv7.2 and Kv7.3 potassium channels.The fre-quency of EEG spiking and sharp-wave issuance was significantly higher and epileptic symptoms were exac-erbated.Conclusion REST/NRSF in mouse hipp-ocampal brain regions is involved in epileptic disease development through transcriptional regulation of Kv7.2 and Kv7.3 potassium channels.

Activation of nuclear factor erythroid 2-related factor 2(Nrf2)by Kelch-like ECH-associated protein 1(Keap1)alkylation plays a central role in anti-inflammatory therapy.However,activators of Nrf2 through alkylation of Keap1-Kelch domain have not been identified.Deoxynyboquinone(DNQ)is a natural small molecule discovered from marine actinomycetes.The current study was designed to investigate the anti-inflammatory effects and molecular mechanisms of DNQ via alkylation of Keap1.DNQ exhibited signif-icant anti-inflammatory properties both in vitro and in vivo.The pharmacophore responsible for the anti-inflammatory properties of DNQ was determined to be the α,β-unsaturated amides moieties by a chemical reaction between DNQ and N-acetylcysteine.DNQ exerted anti-inflammatory effects through activation of Nrf2/ARE pathway.Keap1 was demonstrated to be the direct target of DNQ and bound with DNQ through conjugate addition reaction involving alkylation.The specific alkylation site of DNQ on Keap1 for Nrf2 activation was elucidated with a synthesized probe in conjunction with liquid chromatography-tandem mass spectrometry.DNQ triggered the ubiquitination and subsequent degra-dation of Keap1 by alkylation of the cysteine residue 489(Cys489)on Keap1-Kelch domain,ultimately enabling the activation of Nrf2.Our findings revealed that DNQ exhibited potent anti-inflammatory capacity through α,β-unsaturated amides moieties active group which specifically activated Nrf2 signal pathway via alkylation/ubiquitination of Keap1-Kelch domain,suggesting the potential values of targeting Cys489 on Keap1-Kelch domain by DNQ-like small molecules in inflammatory therapies.

AIM:To investigate the molecular mechanism underlying ferroptosis induced by celastrol(Cel)in huamn pancreatic cancer cell line PANC-1.METHODS:The viability of PANC-1 cells was analyzed by MTT assay,and the effects of Cel on cell proliferation were analyzed using EdU and colony formation assays.Flow cytometry and fluores-cence microscopy were used to assess and observe changes in lipid reactive oxygen species(ROS)levels,respectively,while the levels of malondialdehyde(MDA),glutathione(GSH)and Fe2+were measured using specific kits.The protein expression of glutathione peroxidase 4(GPX4)was evaluated by Western blot,and GPX4 ubiquitination was measured by immunoprecipitation.RESULTS:It was found that the viability,proliferation and colony formation in PANC-1 cells de-creased gradually as the concentration of Cel increased.Addition of Cel alone to the cells reduced both cell rounding and viability,while treatment with ferrostatin-1(Fer-1)alone or in combination with Cel had no effect on either cell morpholo-gy or viability.Fluorescence staining of lipid ROS with BODIPYTM 581/591 C11 followed by flow cytometry analysis showed significantly increased levels of green fluorescence indicative of oxidized lipid ROS,which were further increased after treatment of the cells with Cel.Treatment of the cells with both Cel and Fer-1 reduced the green fluorescence and lip-id ROS levels.Treatment with Cel also increased the levels of MDA and Fe2+,relative to the controls,which reducing the levels of GSH,while addition of both Cel and Fer-1 to the cells restored the levels of MDA,Fe2+,and GSH to those of the control group.CONCLUSION:Treatment with Cel reduces the proliferation of pancreatic cancer cells by inducing fer-roptosis through promoting the ubiquitination and degradation of GPX4.