Objective To investigate the effect of neferine(Nef)on the apoptosis of nucleus pulposus cells under abnormal mechanical stretch stress and its related mechanism.Methods Rat nucleus pulposus cells were subjected to cyclic mechanical stretch stress using Flexcell-5000T tension system,and different concentrations of Nef(6.25,12.5,25 μmol/L)and acetylcysteine(NAC,2 mmol/L)were used to intervention cells.CCK-8 was used to detect cell proliferation activity;Hoechst 33258 fluorescence staining was used to observe the morphological changes of the nucleus;Flow cytometry was used to detect the cell apoptosis level;Biochemical kits were used to detect the levels of lactate dehydrogenase(LDH)in the supernatant,malondialdehyde(MDA)and superoxide dismutase(SOD)in the cells;Western blot was used to detect the expression levels of apoptosis-related proteins,including Bax,Bcl-2,apoptotic protease activating factor-1(Apaf-1),Cleaved-caspase 9,Cleaved-caspase 3,as well as extracellular matrix-associated proteins aggrecan cartilage proteoglycan(Aggrecan),collagen type Ⅱ alpha 1 chain(COL2A1),matrix metalloproteinase-3(MMP-3),and antioxidant signaling pathway proteins nuclear factor erythroid 2-related factor 2(Nrf2)and heme oxygenase 1(HO-1).DCFH-DA fluorescent probe was used to detect the level of reactive oxygen species(ROS)in cells.JC-1 staining was used to detect the level of mitochondrial membrane potential in cells.Results The apoptosis rates of nucleus pulposus cells in the control group and the 5%,10%,15%,and 20%tensile strain groups were(6.69±0.37)%,(8.24±0.33)%,(14.45±0.65)%,(30.48±0.78)%,and(43.06±1.32)%,respectively.Compared with the control group,exposure to 5%,10%,15%,and 20%mechanical stretch stress significantly increased the apoptosis rate of nucleus pulposus cells,the level of LDH in the supernatant,and the levels of ROS and MDA within the cells(P<0.05).Additionally,the levels of SOD and mitochondrial membrane potential were significantly reduced(P<0.05).Simultaneously,the protein expression levels of Bax,Apaf-1,Cleaved-caspase 9,and Cleaved-caspase 3 were markedly upregulated(P<0.05),while the expression level of Bcl-2 protein was downregulated(P<0.05).The apoptosis rates of nucleus pulposus cells in the 15%stretch group,15%stretch+6.25 μmol/L Nef,15%stretch+12.5 μmol/L Nef,and 15%stretch+25 μmol/L Nef groups were(29.74±1.03)%,(25.09±0.67)%,(20.57±0.45)%,and(16.06±0.52)%,respectively.Compared with the 15%stretch group,treatment with 6.25,12.5,and 25 μmol/L Nef significantly reduced the apoptosis rate of nucleus pulposus cells,as well as the levels of LDH in the supernatant and intracellular ROS and MDA(P<0.05);Conversely,the SOD activity and mitochondrial membrane potential were significantly enhanced(P<0.05);Additionally,the protein expression levels of Bax,Apaf-1,Cleaved-caspase 9,Cleaved-caspase 3 and MMP-3 were markedly decreased(P<0.05),while the expression level of Bcl-2,Aggrecan,COL2A1,Nrf2 and HO-1 were increased(P<0.05);Notably,the combination of 25 μmol/L Nef and NAC intervention exhibited a more pronounced effect on inhibiting apoptosis in nucleus pulposus cells induced by abnormal mechanical stretch stress(P<0.05).Conclusion Neferine can inhibit the apoptosis of nucleus pulposus cells induced by abnormal mechanical stretch stress,potentially through scavenging ROS and inhibiting mitochondrial apoptosis pathways.

Sj?gren's syndrome(SS)is one of the common rheumatic diseases in clinical practice.Modern medicine commonly uses drugs such as artificial tears,saliva,glucocorticoids,immunosuppressants,and biologics to control the condition.Clinical practice has shown that in addition to modern medical basic treatment,the use of traditional Chinese medicine(TCM)can help improve the clinical efficacy of SS.According to the symptoms and signs of Sj?gren's syndrome in TCM,it is classified as"dryness and obstruction",and the core pathogenesis of the disease is spleen deficiency and deficiency of body fluids.Subsequently,toxic and pathogenic factors gather,leading to the decline of internal organs.The initial causes are spleen damage,unstable barrier,and invasion of pathogenic factors.The core link is spleen dysfunction,insufficient body fluid,and dryness arising from it.Spleen deficiency generates evil,obstruction of qi,and lack of body fluids are the root causes of illness.The main treatment method is the"spleen strengthening method",which treats spleen deficiency,dampness and stagnation,and the body fluid is not distributed.The treatment focuses on strengthening the spleen and qi,supplementing the lungs and generating fluids.Spleen deficiency leads to loss of vitality,blood stasis obstructs blood vessels,and the treatment is to strengthen the spleen,soothe the liver,remove blood stasis,and unblock the orifices.The spleen yang is not vigorous,and qi transformation is impaired.The treatment is to invigorate the spleen and warm the stomach,promote yang circulation,and promote diuresis.

Objective To comprehensively and systematically evaluate the clinical effect of Chuanzhi Tongluo capsule combined with western medical treatment in patients with cerebral infarction using meta-analysis.Methods Domestic databases were searched,including China Knowledge Network,Wanfang,and Wipu databases,and foreign databases were mainly PubMed.The qualified literature was screened according to the inclusion and exclusion criteria,and was evaluated using the software of RevMan5.3 and Stata14.0 after quality assessment.Results Eight randomized controlled trials(RCTs)on the combination of Chuanzhi Tongluo capsule and western medicine were included,with a total of 728 patients with cerebral infarction,among which 365 cases were in the group of Chuanzhi Tongluo capsule combined with western medicine and 363 cases were in the group of western medicine alone.The results were analyzed in terms of the total effective rate,neurological deficit score such as the National Institutes of Health Stroke Scale(NIHSS),daily living scores,such as the Barthel Index and Activities of Daily Living(ADL)scores and inflammation level(Inflammatory mediator IL-6,ultrasensitive C-reactive protein)as the outcome indexes.Meta-analysis results showed that the total effective rate[95%CI,1.18(1.12,1.25),P<0.00001],neurological function impairment score[95%CI,-1.37(-2.08,-0.67),P=0.0001],daily life score[95%,0.88(0.51,1.25),P<0.00001],and inflammation level[95%CI,-0.75(-0.96,-0.54),P<0.00001]were better than those of the control group and showed significant differences.The incidence of adverse events[95%CI,0.89(0.37,2.15),P=0.80],and the difference was not statistically significant with that of western medicine treatment alone.Conclusion Compared with the control group which only used western medicine,the treatment effect of Chuanzhi Tongluo capsule combined with western medicine in the treatment of cerebral infarction is more significant,and the combined treatment can improve the neurological function of the patients,enhance the quality of daily life,and reduce the level of inflammation more effectively.

The identification of constitutional types in traditional Chinese medicine(TCM)is a crucial component in realizing the concept of"treatment before the onset of disease".In recent years,the"Constitution in Chinese Medicine Questionnaire"has been employed as a standardized tool and has achieved notable success in the field of TCM constitution identification.However,there remains scope for further enhancement of its validity and applicability.Techniques for identifying TCM constitutions based on multidimensional phenotypes continue to emerge,yet a universally applicable constitutional determination model has yet to be established.We propose three research and development directions:The self-assessment questionnaires,the non-invasive point-of-care testing models based on TCM constitution phenotype,and the multidimensional phenotype-based constitution identification platforms.By way of an example,we outline specific developmental ideas with a focus on TCM constitution identification technology based on the analysis of exhaled breath.

Objective To comprehensively evaluate the quality difference of Huaihua powder obtained by the hospital's electromagnetic furnace decoction method and automatic decoction machine decoction method.Methods Five methods(induction cooker decoction(once decoction,twice decoction),normal pressure decoction(once decoction,twice decoction),and pressure decoction)were used to prepare the decoction.The HPLC method used a Shiseido CAPCELL PAK-C18(250 mm×4.6 mm,5 μm)column,with a UV detection wavelength of 254 nm,mobile phase of acetonitrile 0.1%phosphoric acid water,column temperature of 30℃,flow rate of 1.0 mL/min,and detection of index components including rutin,naringin,quercitrin,neohesperidin,quercetin,and menthone.The extraction rate of the index components was calculated and the cream yield of each decoction was detected.The evaluation of quality differences is based on the comprehensive score of cream yield(40%)and indicator components(60%).Results The HPLC fingerprint similarity of Huaihua powder prepared by 5 decoction methods was greater than 99%.There was no significant difference(P>0.05)in the yield of ointment and the content of indicator components between the induction cooker one decoction and the induction cooker twice decoction methods.The yield of ointment and the extraction rate of indicator components were significantly higher in twice normal pressure decoction using a decoction machine than once normal pressure decoction using a decoction machine(P<0.05).The index component score of machine under pressure decoction once was the highest,but the comprehensive score of twice normal pressure decoction was the highest,with a combined score of 92.13 and 89.58,respectively,far higher than the 36.01,18.79,and 15.60 of the other three decoction methods.Classify the decoction into three categories through cluster analysis.Orthogonal partial least squares-discriminant analysis was used to screen for rutin,naringin and neohesperidin as differential biomarkers.Conclusion The quality of Huaihua powder decocted by a decoction machine is significantly better than that by an induction cooker,and the twice decocted at normal pressure by the decoction machine are significantly better than those decocted at normal pressure once.Decoction under pressure once may be a better choice.

Objective Epigenetics,which changes the expression of DNA or protein by chemical modification,affects the regulation of genes and plays an important role in the interaction between genetic and environmental factors in diseases.This review focuses on the development trend and research hotspots of epigenetic modification in ulcerative colitis from 2003 to 2023.Methods Literature related to epigenetic modification in UC was obtained from the Web of Science Core Collection database,and bibliometric analyses and visualization were performed using VOSviewer,CiteSpace and R software.Results 1359 papers published from 2003-2023 were retrieved for inclusion in the analysis.The United States dominated the field of epigenetic modification,with the University of Chicago being the most published and cited institution,and Wu Feng of the Meyerhoff Center for Inflammatory Bowel Disease being the most cited author.The journals Inflammatory Bowel Diseases and Gastroenterology have high influence,screening for high-frequency keywords related to UC epigenetic modifications,including microRNA,inflammation,biomarker,azathioprine,DNA methylation,long noncoding RNA,circular RNA,received wide attention.Conclusion Epigenetics continues to evolve in the field of UC,providing important avenues in elucidating the pathogenesis of UC,in elucidating the interactions between genetics and the environment,and in giving new ideas for simplifying diagnosis,improving inflammation,and developing new drugs.

Objective To construct a diagnostic model for phlegm-dampness syndrome in hypertension based on multimodal feature fusion.Methods Clinical text information(physiological/lifestyle data,symptoms,pulse characteristics)and tongue image data were collected from 261 hypertension patients.For clinical text data,statistical analyses,including ANOVA,Mann-Whitney U test,and Chi-square test,were performed to identify significant features(P<0.05),which were incorporated into a clinical text-based diagnosis model(CTDM)using a multilayer perceptron algorithm.For tongue images,a tongue image-based diagnosis model(TIDM)was constructed based on channel attention mechanisms and residual networks.A multimodal diagnostic model(MDM)was built by fusing clinical text and tongue image features using a feature concatenation method.The diagnostic performance of each model was evaluated using five-fold cross-validation with the area under the receiver operating characteristic curve(AUC),accuracy,specificity,and sensitivity.Results Seven clinical text features,including physiological/lifestyle factors(disease duration,body mass index),symptoms(chest tightness,loss of appetite,excessive phlegm),and pulse characteristics(slippery pulse,damp pulse),were identified as risk factors for phlegm-dampness syndrome in hypertension.The AUC of the CTDM was 0.831±0.021,the AUC of the TIDM was 0.878±0.035,and the MDM achieved an AUC of 0.972±0.015.Conclusion The multimodal diagnostic model that integrates clinical text and tongue image features demonstrates high diagnostic accuracy and provides valuable guidance for AI-assisted diagnosis of phlegm-dampness syndrome in hypertension.

Professor Wei Sun has dedicated extensive research into the prevention and treatment of chronic kidney disease(CKD)using traditional Chinese medicine.His clinical observations identified"kidney deficiency,dampness,and blood stasis"as the core pathogenesis of CKD.Consequently,he formulated the eight-character treatment principle"nourish the kidney,clear dampness,regulate collaterals and eliminate turbidity".He advocated for a comprehensive approach combining"treatment,management,and nurturing"to slow CKD progression,delay dialysis initiation,plan for dialysis effectively,minimize ultrafiltration during dialysis,and reduce complications.These methods aimed to prolong patient survival and enhance their quality of life,achieving a state where patients were"ill but not debilitated,debilitated but gradual",thus allowing better integration into work and daily life.

Objective To investigate the effects of saikosaponin D(SSD)on the growth of esophageal squamous cell carcinoma(ESCC)Eca109 cells and the hypoxia-inducible transcription factor-2α(HIF-2α)/nuclear factor-κB p65(NF-κB p65)pathway.Methods Different concentrations of SSD(0,3,6,9,12,15,18,21,24 μmol·L-1)were used to treat Het-1a normal esophageal squamous epithelial cells and Eca109 ESCC cells to screen the experimental concentration of SSD.Eca109 cells were separated into control(NC)group,HIF-2α inhibitor(PT2385)group,SSD low and high concentration(SSD-L,SSD-H)groups,SSD-H+empty lentivirus(LV-NC)group,and SSD-H+HIF-2α recombinant lentivirus(LV-HIF-2α)group.MTT assay was applied to detect cell viability.Flow cytometry was applied to detect cell apoptosis.ELISA was applied to detect the levels of TNF-α,IL-1β,and IL-8 in cell culture media.Western blot was applied to detect cell proliferation,apoptosis,and protein expression related to the HIF-2α/NF-κB p65 pathway.The Eca109 cell tumor bearing nude mouse model was established to verify the effect of SSD on the in vivo growth of Eca109 cells.Results 6 and 12 μmol·L-1 SSD were selected as experimental concentrations.Compared with the NC group,the cell viability,TNF-α,IL-1β,IL-8 levels in cell culture medium,PCNA,Bcl-2,HIF-2α,p-NF-κB p65/NF-κB p65 protein levels,subcutaneous transplant tumor volume and weight in nude mice,serum TNF-α,IL-1β,IL-8 levels,and positive expression of HIF-2α and NF-κB p65 proteins in the PT2385 group,SSD-L group,and SSD-H group were lower,the apoptosis rate,Bax,and Cleaved-caspase3 protein levels were higher(P<0.05 or P<0.01).Overexpression of HIF-2α prominently weakened the inhibitory effects of SSD on the proliferation of Eca109 cells and the growth of subcutaneous transplanted tumors in nude mice(P<0.05 or P<0.01).Conclusion SSD can inhibit the proliferation of Eca109 cells,induce cell apoptosis,and suppress the growth of subcutaneous transplanted tumors of Eca109 cells in nude mice.Its effect may be related to the inhibition of the activation of the HIF-2α/NF-κB p65 pathway.

Objective By observing the regulatory effect of Strengthening Spleen and Eliminating Cancer Formula on N6-methyladenosine(m6A)methyltransferase,To explore the effect of Strengthening Spleen and Eliminating Cancer Formula on inhibiting the IRX5 m6A level in colorectal cancer(CRC),the regulatory effect on N6-methyladenosine(m6A)methyltransferase was observed.Methods Clinically,m6A hypermethylated genes in colorectal cancer was analyzed by m6A sequencing of pathological tissues from five CRC patients after radical surgery,looking for protein detection indexes for validation.23 BALB/c nude mice were selected and injected with HCT116 cells to establish a nude-mouse transplantation model of human colorectal cancer.They were divided into Model group,Western medicine group(5-fluorouracil group),Chinese medicine group(Jianpi Xiaoai Formula low-dose group,Jianpi Xiaoai Formula high-dose group),with 6 rats in each group,5 rats in control group.The tumor volume of all groups was compared.The overall methylation level of m6A was detected by colorimetric method.The protein expression levels of METTL3,METTL14,and WTAP,in tumor were detected by Western blot.The SRAMP website was used to predict the m6A sites of IRX5.HCT116 cells were treated with oe-NC,oe-METTL3,sh-NC,and sh-METTL3.The expression of IRX5 protein was detected by Western blot.HCT116 cell line was treated with Jianpi Xiaoai Formula drug-containing serum,and transfected with oe-METTL3 and oe-IRX5.The group was set as followed:control group,Jianpi Xiaoai Formula drug-containing serum group,Jianpi Xiaoai Formula drug-containing serum group+oe-NC,Jianpi Xiaoai Formula drug-containing serum group+oe-METTL3,Jianpi Xiaoai Formula drug-containing serum group+oe-IRX5,cell cloning experiment and Transwell experiment were performed to detect cell proliferation,migration and invasion ability of each group.The protein expression levels of METTL3 and IRX5 were detected by Western blot.Results The results of m6A sequencing of genes showed that the m6A methylation level increased in patients with CRC,and the m6A methylation levels of SOX1 and IRX5 were significantly elevated.Compared with the model group,the tumor volume of Jianpi Xiaoyou Formula high-dose group,low-dose group and 5-Fu group decreased significantly(P<0.01),and the tumor inhibition effect was more obvious with the increase of Jianpi Xiaoai Formula concentration(P<0.01).The methylation level of m6A in Jianpi Xiaoai Formula high dose group,low dose group and 5-Fu group decreased significantly(P<0.01).The SRAMP website predicted that IRX5 contained multiple m6A sites.Overexpression of METTL3 promoted the expression of IRX5 protein(P<0.001),while knockdown of METTL3 inhibited the expression of IRX5 protein(P<0.001).The drug-containing serum of Jianpi Xiaoai Formula could inhibit the protein expression of METTL3 and IRX5(P<0.05)and inhibit the proliferation,migration and invasion of HCT116(P<0.01).Overexpression of METTL3 and IRX5 reversed the inhibitory effect of Jianpi Xiaoai Formula on HCT116 evil phenotype(P<0.01).Conclusion Jianpi Xiaoai Formula may inhibit METTL3 expression mediated IRX5 low expression to inhibit the progression of colorectal cancer.