AIM To optimize the extraction process for total polyphenols from Conioselinum vaginatu(Spreng.)Thell.,make component analysis,and evaluate their anti-oxidant,hypoglycemic activities.METHODS The effects of ultrasound,enzymatic hydrolysis,acid hydrolysis,alcohol extraction and hydrolysis processes on the extraction quantity of total polyphenols were investigated,respectively.With extraction temperature,extraction time,ethanol concetration and liquid-solid ratio as influencing factors,extraction quantity of total polyphenols as an evaluation index,the extraction process was optimized by response surface method.HPLC was adopted in the identification of polyphenolic composition and determination of their contents.Subsequently,total polyphenols' scavenging capacities on DPPH,ABTS,OH free radicals,total reducing power and inhibitory capacity on α-glucosidase were determined.RESULTS The highest extraction quantity of total polyphenols was observable when extraction process was employed.The optimal conditions were determined to be 62 ℃ for extraction temperature,54 min for extraction time,69％for ethanol concentration,and 50∶1 for liquid-solid ratio,the extraction quantity of total polyphenols was(9.51±0.2)mg GAE/g.Seven constituents existed in C.vaginatu,among which ferulic acid demonstrated the highest content,followed by that of myricetin,while D-tryptophan content was the lowest.At the concentration of 7.61 mg/L,total polyphenols displayed the scavenging rates on DPPH,ABTS,OH free radicals of 80.70％,85.97％,28.60％,total reducing power of 0.22,and inhibition rate on α-glucosidase of 77.23％,respectively.CONCLUSION This stable and reliable method can be used for the extraction of total polyphenols from C.vaginatum with strong anti-oxidant,hypoglycemic activities.

Objective To investigate the effect of G9a histone methyltransferase inhibitor BIX01294 on the proliferation of vascular smooth muscle cell(VSMC)and its underlying mechanism.Methods Twelve SD rats were divided into the control group and the treatment group,with 6 rats in each group.The rats were anesthetized by intraperitoneal injection with sodium pentobarbital,then exposed the common carotid artery and the internal and external carotid artery after disinfection and skin preparation,ligated the distal end of the internal and external jugular veins,clamped the proximal end of the common carotid artery,cut the external carotid artery,inserted the balloon catheter,blocked the blood flow by compression,and ligated the proximal end of the external carotid artery after withdrawing the balloon.The common carotid artery clamping state of rats in the treatment group was maintained after withdrawing the balloon,then restored the blood flow after perfusion with 100 μmol/L BIX01294 solution for 30 seconds;rats in the control group were restored the blood flow after perfusion with PBS for 30 seconds.VSMC were divided into the normal group and the BIX01294 groups.Cells in the normal group were cultured in low glucose medium containing 2%fetal bovine serum,and cells in the BIX01294 groups were co-treated with 2.5,5.0,7.5 and 10.0 μmol/L BIX01294 on the basis of the normal group,respectively.CCK-8 assay and EDU assay were used to detect the cell activity and proliferative ability respectively,to screen appropriate concentration of BIX01294.The cell apoptosis was detected by TUNEL assay between the normal group and the BIX01294 group,and the control group and the treatment group.Western blot was used to detect the expression levels of autophagy-and apoptosis-related proteins in the normal group and BIX01294 group.Results Compared with 0 μmol/L,the activity and proliferative ability of VSMC decreased in a concentration-dependent manner after treatment with BIX01294 at different concentrations(P<0.05).Compared with the normal group,the apoptosis level of VSMC in the BIX01294 group was increased(P<0.05),the expression of VSMC autophagy-and apoptosis-related proteins of LC3Ⅰ/Ⅱ and Bax were up-regulated(P<0.05),and the expression of Bcl-2 and p62 proteins were down-regulated(P<0.05).In vivo test results showed that BIX01294 local perfusion aggravated the apoptosis of carotid VSMC.Conclusion BIX01294 activates VSMC autophagy and apoptosis and inhibits its proliferation.

Osteosarcoma(OS) is the most common primary bone malignancy with a high propensity for local infiltration and metastasis.c-Jun N-terminal kinases(JNK)is an extremely im-portant member of the mitogen-activated protein kinase(MAPK)family,and the JNK signaling pathway has been shown to be in-volved in regulating OS development.In this paper,we review the research progress on the JNK signaling pathway regulating biological behaviors such as proliferation,migration,invasion,an-giogenesis,autophagy,apoptosis and pyroptosis of OS,as well as the oxidative stress and non-coding RNA regulation of OS through the JNK signaling pathway,to further explore the intrin-sic regulatory mechanisms of the JNK signaling pathway,so as to provide a new way of thinking in searching for the treatment of OS.

p38 mitogen-activated protein kinases(p38 MAPK)are involved in the regulation of osteosarcoma(OS)development.Therefore,this paper reviews the current research status of p38 MAPK signaling pathway in OS,mainly including the p38 MAPK signaling pathway regulates the biological behaviors of OS such as proliferation,migration,invasion,apoptosis,autophagy,iron death,angiogenesis and epithelial-mesenchymal transition(EMT),and non-coding RNAs and oxidative stress are involved in the development of OS through the modulation of p38 MAPK signaling pathway.reviewed to provide new ideas for finding the treatment of OS.

Objective To perform the identification at the subspecies-level of Mycobacterium abscessus(M.abscessus)and analyze its characteristics based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS).Methods Bi-otyper software was used to construct the predicted peak spectrum of M.abscessus subsp.abscessus and M.abscessus subsp.massiliense.The predicted peak spectrum was constructed with expected maximum peak value number of 70 and peak frequency of 100％in the ex-perimental group and control group,respectively.A blind test was performed on 31 strains of M.abscessus that were not used to con-struct predictive peak spectra to evaluate the identification efficiency of predictive peak spectra.FlexAnalysis software was used to sum-marize and analyze the list of mass spectral peak value of M.abscessus,and screen the specific peaks in mass spectra of different sub-species of M.abscessus.The principal component analysis(PCA)algorithm was used to perform the cluster analysis for the data from mass spectrometry of M.abscessus,and explore the feasibility of PCA clustering in distinguishing the subspecies of M.abscessus.Results In the experimental group,96.8％(30/31)of the strains were correctly identified,and one strain of M.abscessus subsp.massiliense with rough colony form was mistakenly identified as M.abscessus subsp.abscessus.In control group,77.4％(24/31)of the strains were correctly identified,but 7 strains of M.abscessus subsp.massiliense were incorrectly identified or unable to be identified.The identification efficiency in the experimental group was significantly better than that in the control group with statistical difference(X2=5.167,P=0.026).M.abscessus subsp.abscessus exhibited three specific peaks(m/z 4 001.67,4 386.81 and 4 963.17),and M.abscessus subsp.massiliense also exhibited three specific peaks(m/z 4 950.48,4 381.78 and 5 214.90).In the PCA 3D scatter plot,the data points of M.abscessus subsp.abscessus and M.abscessus subsp.massiliense were relatively dispersed without obvious clus-tering.The PC A dendrograph could be divided into six branches in which only four branches were composed of a single subspecies.The minimum level value of distance between M.abscessus subsp.abscessus and M.abscessus subsp.massiliense was about 0.1.Conclusion The predicted peak spectrum based on MALDI-TOF MS with the expected maximum peak number of 70 could accurately identify M.abscessus at the subspecies level.The specific peak of mass spectrometry method in this study should be feasible to distinguish the subspecies of M.abscessus subsp.abscessus and the subspecies of M.abscessus subsp.Massiliense,but PCA cluster analysis cannot be used as a means to distinguish M.abscessus subsp.abscessus from M.abscessus subsp.massiliense.

p38 mitogen-activated protein kinases(p38 MAPK)are involved in the regulation of osteosarcoma(OS)development.Therefore,this paper reviews the current research status of p38 MAPK signaling pathway in OS,mainly including the p38 MAPK signaling pathway regulates the biological behaviors of OS such as proliferation,migration,invasion,apoptosis,autophagy,iron death,angiogenesis and epithelial-mesenchymal transition(EMT),and non-coding RNAs and oxidative stress are involved in the development of OS through the modulation of p38 MAPK signaling pathway.reviewed to provide new ideas for finding the treatment of OS.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.

Objective To investigate the effect of G9a histone methyltransferase inhibitor BIX01294 on the proliferation of vascular smooth muscle cell(VSMC)and its underlying mechanism.Methods Twelve SD rats were divided into the control group and the treatment group,with 6 rats in each group.The rats were anesthetized by intraperitoneal injection with sodium pentobarbital,then exposed the common carotid artery and the internal and external carotid artery after disinfection and skin preparation,ligated the distal end of the internal and external jugular veins,clamped the proximal end of the common carotid artery,cut the external carotid artery,inserted the balloon catheter,blocked the blood flow by compression,and ligated the proximal end of the external carotid artery after withdrawing the balloon.The common carotid artery clamping state of rats in the treatment group was maintained after withdrawing the balloon,then restored the blood flow after perfusion with 100 μmol/L BIX01294 solution for 30 seconds;rats in the control group were restored the blood flow after perfusion with PBS for 30 seconds.VSMC were divided into the normal group and the BIX01294 groups.Cells in the normal group were cultured in low glucose medium containing 2%fetal bovine serum,and cells in the BIX01294 groups were co-treated with 2.5,5.0,7.5 and 10.0 μmol/L BIX01294 on the basis of the normal group,respectively.CCK-8 assay and EDU assay were used to detect the cell activity and proliferative ability respectively,to screen appropriate concentration of BIX01294.The cell apoptosis was detected by TUNEL assay between the normal group and the BIX01294 group,and the control group and the treatment group.Western blot was used to detect the expression levels of autophagy-and apoptosis-related proteins in the normal group and BIX01294 group.Results Compared with 0 μmol/L,the activity and proliferative ability of VSMC decreased in a concentration-dependent manner after treatment with BIX01294 at different concentrations(P<0.05).Compared with the normal group,the apoptosis level of VSMC in the BIX01294 group was increased(P<0.05),the expression of VSMC autophagy-and apoptosis-related proteins of LC3Ⅰ/Ⅱ and Bax were up-regulated(P<0.05),and the expression of Bcl-2 and p62 proteins were down-regulated(P<0.05).In vivo test results showed that BIX01294 local perfusion aggravated the apoptosis of carotid VSMC.Conclusion BIX01294 activates VSMC autophagy and apoptosis and inhibits its proliferation.

Neuropathic pain, a debilitating condition caused by dysfunction of the somatosensory nervous system, remains difficult to treat due to limited understanding of its molecular mechanisms. Bioinformatics analysis identified cerebellin 2 (CBLN2) as highly enriched in human and murine proprioceptive and nociceptive neurons. We found that CBLN2 expression is persistently upregulated in dorsal root ganglia (DRG) following spinal nerve ligation (SNL) in mice. In addition, transcription factor SOX11 binds to 12 cis-regulatory elements within the Cbln2 promoter to enhance its transcription. SNL also induced SOX11 upregulation, with SOX11 and CBLN2 co-localized in nociceptive neurons. The siRNA-mediated knockdown of Sox11 or Cbln2 attenuated SNL-induced mechanical allodynia and thermal hyperalgesia. High-throughput sequencing of DRG following intrathecal injection of CBLN2 revealed widespread gene expression changes, including upregulation of numerous NF-κB downstream targets. Consistently, CBLN2 activated NF-κB signaling, and inhibition with pyrrolidine dithiocarbamate reduced CBLN2-induced pain hypersensitivity, proinflammatory cytokines and chemokines production, and neuronal hyperexcitability. Together, these findings identified the SOX11/CBLN2/NF-κB axis as a critical mediator of neuropathic pain and a promising target for therapeutic intervention.
Animals ; Neuralgia/metabolism* ; Ganglia, Spinal/metabolism* ; Up-Regulation ; Mice ; NF-kappa B/metabolism* ; SOXC Transcription Factors/genetics* ; Male ; Neuroinflammatory Diseases/metabolism* ; Mice, Inbred C57BL ; Nerve Tissue Proteins/genetics* ; Hyperalgesia/metabolism* ; Signal Transduction ; Spinal Nerves

Objective To perform the identification at the subspecies-level of Mycobacterium abscessus(M.abscessus)and analyze its characteristics based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS).Methods Bi-otyper software was used to construct the predicted peak spectrum of M.abscessus subsp.abscessus and M.abscessus subsp.massiliense.The predicted peak spectrum was constructed with expected maximum peak value number of 70 and peak frequency of 100％in the ex-perimental group and control group,respectively.A blind test was performed on 31 strains of M.abscessus that were not used to con-struct predictive peak spectra to evaluate the identification efficiency of predictive peak spectra.FlexAnalysis software was used to sum-marize and analyze the list of mass spectral peak value of M.abscessus,and screen the specific peaks in mass spectra of different sub-species of M.abscessus.The principal component analysis(PCA)algorithm was used to perform the cluster analysis for the data from mass spectrometry of M.abscessus,and explore the feasibility of PCA clustering in distinguishing the subspecies of M.abscessus.Results In the experimental group,96.8％(30/31)of the strains were correctly identified,and one strain of M.abscessus subsp.massiliense with rough colony form was mistakenly identified as M.abscessus subsp.abscessus.In control group,77.4％(24/31)of the strains were correctly identified,but 7 strains of M.abscessus subsp.massiliense were incorrectly identified or unable to be identified.The identification efficiency in the experimental group was significantly better than that in the control group with statistical difference(X2=5.167,P=0.026).M.abscessus subsp.abscessus exhibited three specific peaks(m/z 4 001.67,4 386.81 and 4 963.17),and M.abscessus subsp.massiliense also exhibited three specific peaks(m/z 4 950.48,4 381.78 and 5 214.90).In the PCA 3D scatter plot,the data points of M.abscessus subsp.abscessus and M.abscessus subsp.massiliense were relatively dispersed without obvious clus-tering.The PC A dendrograph could be divided into six branches in which only four branches were composed of a single subspecies.The minimum level value of distance between M.abscessus subsp.abscessus and M.abscessus subsp.massiliense was about 0.1.Conclusion The predicted peak spectrum based on MALDI-TOF MS with the expected maximum peak number of 70 could accurately identify M.abscessus at the subspecies level.The specific peak of mass spectrometry method in this study should be feasible to distinguish the subspecies of M.abscessus subsp.abscessus and the subspecies of M.abscessus subsp.Massiliense,but PCA cluster analysis cannot be used as a means to distinguish M.abscessus subsp.abscessus from M.abscessus subsp.massiliense.