As a non-invasive biomarker, circulating tumor DNA (ctDNA) can sensitively identify minimal residual disease (MRD), offering a novel approach for prognosis prediction and efficacy evaluation in patients with colorectal cancer liver metastasis (CRLM), thereby aiding in the formulation of personalized treatment strategies. This article summarized the progress in the application of ctDNA detection for MRD in CRLM and provided insights into its future directions.

Objective·To comprehensively evaluate the efficiency of different kits and methods for DNA and RNA extraction from blood samples.Methods·A total of 145 blood samples were collected,including those from patients with Alzheimer's disease(20 cases),fibrosis(5 cases),colorectal cancer(108 cases),and healthy individuals(12 cases).A column-based kit(Kit A)and a nucleic acid extraction instrument were used to extract genomic DNA(gDNA)from leukocytes in the blood.Cell-free DNA(cfDNA)and cell-free RNA(cfRNA)in plasma were extracted using five different kits(Kit B?F),which employed either column-based(Kit B,E)or magnetic bead-based methods(Kit C,D,F).The extraction process of Kit B was optimized by increasing the plasma sample volume and extending the elution incubation time.Furthermore,this protocol was applied to extracting cfDNA from plasma samples of 100 colorectal cancer patients.Quantitative real-time PCR(qPCR)was used to quantify the extracted DNA and RNA,and the molecular yields were compared to evaluate the extraction efficiency.A comprehensive assessment was conducted,considering factors such as cost and operation time.Results·In gDNA extraction,although the the operation time was shortened by using the nucleic acid extraction instrument,the median number of DNA molecules extracted using Kit A(column-based method)was 25.36-fold higher than that obtained with the instrument(P<0.05).For cfDNA extraction,while the overall efficiency of the three kits(Kit B?D)was similar,Kit B(column-based method)showed superior performance in low-concentration samples,with average DNA yields 4.24-fold and 1.18-fold higher than those of Kit D and Kit C(both magnetic bead-based).Optimization of Kit B's extraction protocol further improved cfDNA yield.When comparing three samples,the cfDNA yields from larger plasma input volumes was 3.98-fold,2.38-fold,and 3.82-fold higher than those from smaller input volumes,respectively.The results of cfDNA extraction from 100 colorectal cancer patients indicated that this extraction protocol reliably extracted sufficient amounts of cfDNA from clinical samples.For cfRNA extraction,Kit E(column-based method)was widely recommended due to its high efficiency,convenience,and cost-effectiveness.The median RNA content extracted using Kit E was 5.01-fold higher than that of Kit F(magnetic bead-based method).Lastly,a comparison of the copy numbers of cfDNA and cfRNA in plasma revealed that the average copy number of cfRNA per milliliter of plasma was 27.65-fold higher than that of cfDNA.Conclusion·Kit A,Kit B,and Kit E show outstanding performance in leukocyte gDNA extraction,plasma cfDNA extraction,and plasma cfRNA extraction,respectively.However,although Kit E has advantages in extraction efficiency and cost,its safety requires further evaluation.

Objective·To comprehensively evaluate the efficiency of different kits and methods for DNA and RNA extraction from blood samples.Methods·A total of 145 blood samples were collected,including those from patients with Alzheimer's disease(20 cases),fibrosis(5 cases),colorectal cancer(108 cases),and healthy individuals(12 cases).A column-based kit(Kit A)and a nucleic acid extraction instrument were used to extract genomic DNA(gDNA)from leukocytes in the blood.Cell-free DNA(cfDNA)and cell-free RNA(cfRNA)in plasma were extracted using five different kits(Kit B?F),which employed either column-based(Kit B,E)or magnetic bead-based methods(Kit C,D,F).The extraction process of Kit B was optimized by increasing the plasma sample volume and extending the elution incubation time.Furthermore,this protocol was applied to extracting cfDNA from plasma samples of 100 colorectal cancer patients.Quantitative real-time PCR(qPCR)was used to quantify the extracted DNA and RNA,and the molecular yields were compared to evaluate the extraction efficiency.A comprehensive assessment was conducted,considering factors such as cost and operation time.Results·In gDNA extraction,although the the operation time was shortened by using the nucleic acid extraction instrument,the median number of DNA molecules extracted using Kit A(column-based method)was 25.36-fold higher than that obtained with the instrument(P<0.05).For cfDNA extraction,while the overall efficiency of the three kits(Kit B?D)was similar,Kit B(column-based method)showed superior performance in low-concentration samples,with average DNA yields 4.24-fold and 1.18-fold higher than those of Kit D and Kit C(both magnetic bead-based).Optimization of Kit B's extraction protocol further improved cfDNA yield.When comparing three samples,the cfDNA yields from larger plasma input volumes was 3.98-fold,2.38-fold,and 3.82-fold higher than those from smaller input volumes,respectively.The results of cfDNA extraction from 100 colorectal cancer patients indicated that this extraction protocol reliably extracted sufficient amounts of cfDNA from clinical samples.For cfRNA extraction,Kit E(column-based method)was widely recommended due to its high efficiency,convenience,and cost-effectiveness.The median RNA content extracted using Kit E was 5.01-fold higher than that of Kit F(magnetic bead-based method).Lastly,a comparison of the copy numbers of cfDNA and cfRNA in plasma revealed that the average copy number of cfRNA per milliliter of plasma was 27.65-fold higher than that of cfDNA.Conclusion·Kit A,Kit B,and Kit E show outstanding performance in leukocyte gDNA extraction,plasma cfDNA extraction,and plasma cfRNA extraction,respectively.However,although Kit E has advantages in extraction efficiency and cost,its safety requires further evaluation.

Objective To explore the effect of maternal exercise on blood pressure,cardiac pheno-type and susceptibility to myocardial is chemia/reperfusion(MI/R)injury in adult male offsprings of mice.Methods Pregnant mice were randomly divided into a sedentary group(p-Ctr)and an exercise group(p-EX),each of 12.The exercise group underwent daily 60-minutenon-weight bearing swim-ming from gestational day(GD)1 to 18,6 days a week.Then,their male offsprings at 3 months of age(3M)were selected as the research subjects,namely the Ctr-3M group and the EX-3M group,with 12 mice in each group.The MI/R model was established by ligation of the anterior descending branch of the left coronary artery(LAD)surgery(30 min of ischemia,24 h of reperfusion).Then,the systolic blood pressure(SBP),diastolic blood pressure(DBP),and mean arterial pressure(MAP)were monitored non-invasively via tail artery blood pressure measurement,while the cardiac function was detected by using the small animal ultrasound.Moreover,the cardiac morphology and col-lagen volume fraction(CVF)of myocardium was observed by HE staining and Masson staining,respec-tively.Meanwhile,the cross-sectional area(CSA)of myocardial cells and myocardial infarction area(INF/AAR)was measured using WGA staining and Evans Blue-TTC double staining,accordingly.What's more,the apoptosis index(AI)of myocardial cells and serum cardiac troponin I(cTnI)were de-tected by using TUNEL staining and ELISA,respectively,while the expression levels of Bax and Bcl-2 proteins were determined by using Western blotting.Results 1)From GD11 to GD19,the body weight of females in the p-EX group was always significantly lower than that in the p-Ctr group(GD11,GD13;P<0.05;GD12,GD14～GD19;P<0.01),but there was no significant difference in the litter size and abortion rate(P>0.05).2)There were no significant differences between the EX-3M and Ctr-3M groups in the heart weight(HW),body weight(BW)and heart weight to body weight ratio(HW/BW),SBP,DBP,MAP,myocardial CVF,and CSA of male offsprings(P>0.05).After MI/R surgery,in both p-Ctr and p-EX groups,myocardial transverse striations disappeared and cardiomyocytes were disarranged with degeneration and necrosis.Moreover,CVF of the EX-3M group was significantly lower than the Ctr-3M group after MI/R surgery(P<0.05).3)No significant differenc-es were found between the Ctr-3M and EX-3M groups in the average cardiac ejection fraction(EF),fractional shortening(FS),left ventricle internal diameter in diastole(LVIDd),and left ventricle inter-nal diameter in systole(LVIDs)(P>0.05).However,after the MI/R surgery,the postoperative EF and FS(P<0.01),and LVIDs(P<0.05)in the EX-3M group were significantly higher than the Ctr-3M group,without significant difference in LVIDd(P>0.05).4)After MI/R surgery,the average AI,INF/AAR and serum cTnI level of the EX-3M group were significantly lower than the Ctr-3M group(P<0.01 for all).5)The relative expression level of myocardial Bcl-2 protein after MI/R surgery in the EX-3M group was significantly higher than the Ctr-3M group(P<0.05),while that of myocardial Bax protein(P<0.05)and the Bax/Bcl-2 ratio(P<0.01)were significantly lower than the latter.Conclu-sions Prenatal exercise significantly reduces the susceptibility to myocardial ischemia injury in 3-month-old male mice offspring,improves the contractile function of their heart,and attenuates the degree of cardiomyocyte apoptosis and necrosis,exerting a cardioprotective effect in MI/R injury.

ABSTRCT AIM:To verify the effect of polygona-tum polysaccharide(PSP)combined with TGF-β3 in inducing the differentiation of rat tendon-derived stem cells(TDSCs)into chondrocytes by activating the TGF-β3/Smad2 pathway.METHODS:TDSCs were extracted from rat tail tendons using enzyme digestion,purified,passaged,and identified via flow cytometry.TDSCs were treated with different concentrations of PSP,and the optimal growth con-centration was determined using the CCK-8 assay.TDSCs were divided into four groups:PSP,TGF-β3,PSP+TGF-β3,and Control for differentiation induc-tion.Chondrogenic differentiation was evaluated using morphological observations,toluidine blue staining,immunofluorescence staining,and West-ern blot analysis to detect COLⅡ,SOX9,and AGG.Western blot was also used to measure the expres-sion levels of Smad2 and p-Smad2 to evaluate the activation of the TGF-β3/Smad2 pathway after chondrogenic induction.RESULTS:Flow cytometry analysis showed that TDSCs highly expressed CD90 and CD29,while CD11b and CD45 expression was low.The CCK-8 assay indicated that 5 μmol/L PSP was the optimal intervention dose.Toluidine blue staining revealed that the blue staining area in the PSP,PSP+TGF-β3,and TGF-β3 groups was larger compared to the Control group.Immunofluores-cence analysis demonstrated that COLⅡ expression was significantly increased in the PSP,TGF-β3,and PSP+TGF-β3 groups,with the highest expression in the PSP+TGF-β3 group(P＜0.05).Western blot analy-sis showed that the levels of p-Smad2/Smad2,COLⅡ,SOX9,and AGG were elevated in the PSP,TGF-β3,and PSP+TGF-β3 groups,with the highest ex-pression in the PSP+TGF-β3 group(P＜0.05).Compared to the Control group,the TGF-β3 and PSP groups also showed significantly increased expression(P＜0.05).CONCLUSION:PSP promotes the proliferation and chondrogenic differentiation of TDSCs,possibly by activating the TGF-β3/Smad2 pathway.

ABSTRCT AIM:To verify the effect of polygona-tum polysaccharide(PSP)combined with TGF-β3 in inducing the differentiation of rat tendon-derived stem cells(TDSCs)into chondrocytes by activating the TGF-β3/Smad2 pathway.METHODS:TDSCs were extracted from rat tail tendons using enzyme digestion,purified,passaged,and identified via flow cytometry.TDSCs were treated with different concentrations of PSP,and the optimal growth con-centration was determined using the CCK-8 assay.TDSCs were divided into four groups:PSP,TGF-β3,PSP+TGF-β3,and Control for differentiation induc-tion.Chondrogenic differentiation was evaluated using morphological observations,toluidine blue staining,immunofluorescence staining,and West-ern blot analysis to detect COLⅡ,SOX9,and AGG.Western blot was also used to measure the expres-sion levels of Smad2 and p-Smad2 to evaluate the activation of the TGF-β3/Smad2 pathway after chondrogenic induction.RESULTS:Flow cytometry analysis showed that TDSCs highly expressed CD90 and CD29,while CD11b and CD45 expression was low.The CCK-8 assay indicated that 5 μmol/L PSP was the optimal intervention dose.Toluidine blue staining revealed that the blue staining area in the PSP,PSP+TGF-β3,and TGF-β3 groups was larger compared to the Control group.Immunofluores-cence analysis demonstrated that COLⅡ expression was significantly increased in the PSP,TGF-β3,and PSP+TGF-β3 groups,with the highest expression in the PSP+TGF-β3 group(P＜0.05).Western blot analy-sis showed that the levels of p-Smad2/Smad2,COLⅡ,SOX9,and AGG were elevated in the PSP,TGF-β3,and PSP+TGF-β3 groups,with the highest ex-pression in the PSP+TGF-β3 group(P＜0.05).Compared to the Control group,the TGF-β3 and PSP groups also showed significantly increased expression(P＜0.05).CONCLUSION:PSP promotes the proliferation and chondrogenic differentiation of TDSCs,possibly by activating the TGF-β3/Smad2 pathway.

Endometrial cancer (EC) is one of the most common gynecological malignancies, with an increasing incidence rate worldwide. Accurate segmentation of lesion areas in computed tomography (CT) images is a critical step in assisting clinical diagnosis. In this study, we propose a novel deep learning-based segmentation model, termed spatial choice and weight union network (SCWU-Net), which incorporates two newly designed modules: the spatial selection module (SSM) and the combination weight module (CWM). The SSM enhances the model's ability to capture contextual information through deep convolutional blocks, while the CWM, based on joint attention mechanisms, is employed within the skip connections to further boost segmentation performance. By integrating the strengths of both modules into a U-shaped multi-scale architecture, the model achieves precise segmentation of EC lesion regions. Experimental results on a public dataset demonstrate that SCWU-Net achieves a Dice similarity coefficient (DSC) of 82.98%, an intersection over union (IoU) of 78.63%, a precision of 92.36%, and a recall of 84.10%. Its overall performance is significantly outperforming other state-of-the-art models. This study enhances the accuracy of lesion segmentation in EC CT images and holds potential clinical value for the auxiliary diagnosis of endometrial cancer.
Humans ; Endometrial Neoplasms/diagnostic imaging* ; Female ; Tomography, X-Ray Computed/methods* ; Deep Learning ; Algorithms ; Image Processing, Computer-Assisted/methods* ; Neural Networks, Computer

This article introduces Professor LIU Xing's clinical experience in treatment of peripheral facial paralysis at the recovery and sequelae stages with the combination of acupotomy, cupping and herbal medication. Based on the analysis of etiology and pathogenesis of peripheral facial paralysis, Professor LIU believes that "invasion of pathogenic wind to collaterals and obstruction of qi and blood" is crucial. Therefore, the treatment focuses on "dispelling wind and harmonizing blood". The compound therapeutic mode is proposed, with acupotomy, cupping and herbal decoction involved, in which, "three-step sequential method of acupotomy" is predominated. Firstly, in the prone position, five "feng" (wind) points are stimulated in patient, Fengfu (GV16), Fengchi (GB20), Yifeng (TE17), Bingfeng (SI12) and Fengmen (BL12). Secondly, in the lateral position, three-facial points are stimulated (FaceⅠneedle: Yangbai [GB14]-Yuyao [EX-HN4]; Face Ⅱ needle: Sibai [ST2]-Quanliao [SI18]; Face Ⅲ needle: Jiache [ST6]-Dicang [ST4]) to restore the deviated facial muscles. Finally, in the supine, two Dantian points are stimulated on the forehead and chest, respectively (upper Dantian: Yintang [GV24⁺], middle Dantian: Danzhong [CV17]), to regulate qi and blood. As the adjunctive therapies, cupping is used to remove stasis, and herbal decoction is to harmonize the body interior. In view of holistic regulation, the treatment is administered in accordance with the affected meridians, so as to expel wind, remove obstruction in collaterals and regulate qi and blood.
Humans ; Facial Paralysis/drug therapy* ; Drugs, Chinese Herbal/administration & dosage* ; Acupuncture Therapy ; Male ; Female ; Middle Aged ; Adult ; Combined Modality Therapy ; Acupuncture Points ; Cupping Therapy ; Aged ; Young Adult

OBJECTIVES: To explore the active components that mediate the therapeutic effect of Centella asiatica on psoriasis and their therapeutic mechanisms. METHODS: TCMSP, TCMIP, PharmMapper, Swiss Target Prediction, GeneCards, OMIM and TTD databases were searched for the compounds in Centella asiatica and their targets and the disease targets of psoriasis. A drug-active component-target network and the protein-protein interaction network were constructed, and DAVID database was used for pathway enrichment analysis. In a RAW264.7 macrophage model of LPS-induced inflammation, the anti-inflammatory effect of 7.5, 15, 30, and 60 μmol/L quercetin, asiaticoside, and asiatic acid, which were identified as the main active components in Centella asiatica, were tested by measuring cellular production of NO, TNF‑α and IL-6 using Griess method and ELISA and by detecting mRNA expressions of IL-23, IL-17A, TNF-α and IL-6 and protein expressions of p-STAT3 (Tyr705) and p-STAT3 (Ser727) with RT-qPCR and Western blotting. RESULTS: A total of 139 targets of Centella asiatica and 4604 targets of psoriasis were obtained, and among them CASP3, EGFR, PTGS2, and ESR1 were identified as the core targets. KEGG analysis suggested that quercetin, asiaticoside, and asiatic acid in Centella asiatica were involved in cancer and IL-17 and MAPK signaling pathways. In the RAW264.7 macrophage model of inflammation, treatment with quercetin significantly reduced cellular production of NO, TNF‑α and IL-6, and lowered mRNA expressions of IL-23, IL-17A, TNF‑α and IL-6 and protein expressions of p-STAT3 (Tyr705) and p-STAT3 (Ser727). CONCLUSIONS Quercetin, asiaticoside and asiatic acid are the main active components in Centella asiatica to mediate the therapeutic effect against psoriasis, and quercetin in particular is capable of suppressing cellular production of NO, TNF‑α and IL-6 and regulating the IL-23/IL-17A inflammatory axis by mediating STAT3 phosphorylation to inhibit inflammatory response.
Quercetin/pharmacology* ; Psoriasis/metabolism* ; STAT3 Transcription Factor/metabolism* ; Mice ; Animals ; Centella/chemistry* ; Triterpenes/pharmacology* ; Phosphorylation ; Interleukin-17/metabolism* ; Interleukin-23/metabolism* ; RAW 264.7 Cells ; Pentacyclic Triterpenes/pharmacology* ; Macrophages/drug effects* ; Signal Transduction ; Plant Extracts

AIM:To clarify the molecular mechanism by which astragaloside Ⅳ(AS-Ⅳ)suppresses oxida-tive stress and alleviates cerebral ischemia-reperfusion injury(CIRI)via the PTEN-induced kinase 1(PINK1)/parkin mi-tophagy-associated pathway.METHODS:A middle cerebral artery occlusion/reperfusion(MCAO/R)model was estab-lished in Sprague-Dawley rats.The animals were allocated to sham,MCAO/R,AS-Ⅳ,and mitochondrial division inhibi-tor-1(Mdivi-1)treatment groups.The rats in AS-Ⅳ and Mdivi-1 groups were intraperitoneally injected once daily with AS-Ⅳ(20 mg/kg)for 7 d,while those in Midivi-1 group also received intraperitoneal injection of Mdivi-1(1.2 mg·kg-1·d-1).The rats in sham and MCAO/R groups were given equivalent volume of distilled water.Neurological deficits were as-sessed using Zea Longa scoring,infarcted area volumes were measured using TTC staining,and brain tissue pathology was examined using hematoxylin and eosin staining.The levels of malondialdehyde(MDA)and superoxide dismutase(SOD)were assessed by ELISA,while those of reactive oxygen species(ROS)were measured using flow cytometry.The expres-sion levels of PINK1,parkin and microtubule-associated protein 1 light chain 3(LC3)were quantified using Western blot and RT-qPCR.RESULTS:AS-Ⅳ administration significantly alleviated neuronal and mitochondrial damage in MCAO/R model rat brains(P＜0.05),together with significant reductions in the cerebral infarct volume and neurological dysfunc-tion(P＜0.05).Significant increases in PINK1,parkin and LC3 protein and mRNA levels were observed in response to AS-Ⅳ(P＜0.05),SOD activity rose,and ROS and MDA levels declined significantly(P＜0.05).The co-administration of Mdivi-1 abrogated the protective benefits of AS-Ⅳ,inhibited activation of the PINK1/parkin pathway,down-regulated LC3 at the mRNA and protein levels,and significantly increased mitochondrial damage.Mdivi-1 also markedly reduced autophagosome formation and SOD activity level,but increased both ROS and MDA levels,cerebral infarct volume,and the severity of neurological deficits(P＜0.05).CONCLUSION:Astragaloside Ⅳ activates the PINK/parkin-mediated mitophagy pathway,inhibits oxidative stress and alleviates CIRI in rats.