Objective:To improve the quality standards for Portulaca oleracea medicinal materials and decoction pieces.Methods:Fourteen batches of P.oleracea medicinal materials and 20 batches of its decoction pieces from different producing areas across the country were collected.In accordance with the relevant methods in the general chapters of the Chinese Pharmacopoeia(2020 Edition,Volume Ⅰ),the thin-layer chromatography(TLC)identi-fication method was optimized.The contents of impurities,moisture,total ash,acid-insoluble ash,heavy metals and harmful elements,and extracts were determined.Meanwhile,the HPLC feature chromatograms and content determination methods for P.oleracea medicinal materials and decoction pieces were established.Results:The optimized TLC method showed clear spots,good separation effect and reproducibility.The average contents of impurities,moisture,total ash,acid-insoluble ash,and extracts in the 14 batches of P.oleracea medicinal materi-als were 0.01％,8.48％,21.68％,5.33％,and 28.44％respectively.The average contents of impurities,moisture,total ash,acid-insoluble ash,and extracts in the 20 batches of P.oleracea decoction pieces were 0.01％,7.00％,21.09％,3.60％,and 29.63％respectively.The results of the tests for heavy metals and harmful elements showed that the contents of lead,cadmium,arsenic,mercury and copper in 34 batches of samples varied greatly.Moreover,the contents of cadmium,arsenic and copper in some samples exceeded the limit guid-ance values specified in the Pharmacopoeia.Nine common peaks were calibrated in the established HPLC feature chromatogram of P.oleracea,and an HPLC method for simultaneously determining the contents of norepinephrine and dopamine was established.Conclusion:It is recommended to modify the developing solvent for the thin-layer identification of P.oleracea and its proportion to water-saturated n-butanol-acetone-glacial acetic acid-water=4∶1∶1∶1,and change the extraction method of the test sample to ultrasonic extraction for 30 minutes.It is proposed to add the stipulations that the total ash of P.oleracea medicinal materials and decoction pieces should not exceed 25.0％,the acid-insoluble ash should not exceed 6.0％,and the water-soluble extract should not be lower than 20.0％.For P.oleracea decoction pieces,the total ash should not exceed 28.0％,the acid-insoluble ash should not exceed 6.0％,and the water-soluble extract should not be lower than 20.0％.This study provides an experi-mental basis for the improvement of the quality standards of P.oleracea.

Objective:To improve the quality standards for Portulaca oleracea medicinal materials and decoction pieces.Methods:Fourteen batches of P.oleracea medicinal materials and 20 batches of its decoction pieces from different producing areas across the country were collected.In accordance with the relevant methods in the general chapters of the Chinese Pharmacopoeia(2020 Edition,Volume Ⅰ),the thin-layer chromatography(TLC)identi-fication method was optimized.The contents of impurities,moisture,total ash,acid-insoluble ash,heavy metals and harmful elements,and extracts were determined.Meanwhile,the HPLC feature chromatograms and content determination methods for P.oleracea medicinal materials and decoction pieces were established.Results:The optimized TLC method showed clear spots,good separation effect and reproducibility.The average contents of impurities,moisture,total ash,acid-insoluble ash,and extracts in the 14 batches of P.oleracea medicinal materi-als were 0.01％,8.48％,21.68％,5.33％,and 28.44％respectively.The average contents of impurities,moisture,total ash,acid-insoluble ash,and extracts in the 20 batches of P.oleracea decoction pieces were 0.01％,7.00％,21.09％,3.60％,and 29.63％respectively.The results of the tests for heavy metals and harmful elements showed that the contents of lead,cadmium,arsenic,mercury and copper in 34 batches of samples varied greatly.Moreover,the contents of cadmium,arsenic and copper in some samples exceeded the limit guid-ance values specified in the Pharmacopoeia.Nine common peaks were calibrated in the established HPLC feature chromatogram of P.oleracea,and an HPLC method for simultaneously determining the contents of norepinephrine and dopamine was established.Conclusion:It is recommended to modify the developing solvent for the thin-layer identification of P.oleracea and its proportion to water-saturated n-butanol-acetone-glacial acetic acid-water=4∶1∶1∶1,and change the extraction method of the test sample to ultrasonic extraction for 30 minutes.It is proposed to add the stipulations that the total ash of P.oleracea medicinal materials and decoction pieces should not exceed 25.0％,the acid-insoluble ash should not exceed 6.0％,and the water-soluble extract should not be lower than 20.0％.For P.oleracea decoction pieces,the total ash should not exceed 28.0％,the acid-insoluble ash should not exceed 6.0％,and the water-soluble extract should not be lower than 20.0％.This study provides an experi-mental basis for the improvement of the quality standards of P.oleracea.

Objective:To investigate the current self-compassion levels in hematologic malignant tumor patients and analyze the influencing factors, providing a basis for clinical interventions.Methods:A convenience sampling method was used to select 120 patients with hematologic malignant tumors treated at the First Affiliated Hospital of Wenzhou Medical University from January to December 2022. Clinical data, self-compassion levels, social support, and adult attachment status were assessed using general data questionnaires, the Self-Compassion Scale (SCS) , the Perceived Social Support Scale (PSSS) , and the Experiences in Close Relationship Scale-Short Form (ECR-S) . Pearson correlation analysis was performed to analyze the correlation between SCS scores and PSSS, ECR-S scores. Multiple linear regression analysis was conducted to explore the influencing factors of self-compassion levels.Results:A total of 120 questionnaires were distributed, and 114 valid questionnaires were returned, resulting in an effective response rate of 95.00%. The average SCS score of hematologic malignant tumor patients was (80.54±5.80) . Univariate analysis showed that self-compassion levels differed significantly among patients of different ages, education levels, per capita family monthly income, disease duration, and the first diagnosis or not of hematologic malignant tumors ( P<0.01) . Pearson correlation analysis revealed a positive correlation between SCS scores and PSSS scores ( r=0.659, P<0.001) , and a negative correlation with the attachment anxiety and avoidance dimensions of the ECR-S ( r=-0.152, -0.588, P<0.001) . Multiple linear regression analysis showed that education level, the first diagnosis or not, PSSS score, ECR-S attachment anxiety dimension, and ECR-S attachment avoidance dimension scores entered the regression equation. Conclusions:The influencing factors of self-compassion levels in hematologic malignant tumor patients include education level, the first diagnosis or not, social support level, and adult attachment status. Healthcare providers should strengthen psychological support for patients with low education, disease recurrence/treatment failure, and insecure attachment types, while also improving patients' social support levels to enhance their self-compassion status.

BACKGROUND:Extracellular vesicles have received extensive attention in the field of bone defect regeneration and repair in recent years.However,natural extracellular vesicles have deficiencies in sustained controlled release,tissue targeting,and drug loading capacity.Therefore,the introduction of engineering strategies to modify extracellular vesicles to enhance their therapeutic efficacy has become a research hotspot.OBJECTIVE:To review the role and application progress of engineered extracellular vesicles in the regeneration and repair of bone defects.METHODS:PubMed,Web of Science,CNKI,and WanFang databases were searched for relevant articles published in the past fifteen years.The search terms were"engineering,extracellular vesicles,exosomes,bone defect,bone regeneration,bone repair"in Chinese and English.After removal of poorly related,outdated,and duplicate studies by screening,93 articles were finally included for review according to inclusion criteria.RESULTS AND CONCLUSION:(1)Extracellular vesicles are primarily isolated based on their density,size,immunoaffinity,and surface charge.After isolation,extracellular vesicles are characterized using imaging techniques,size-and counting-based techniques,and flow cytometry.(2)Extracellular vesicles stimulate bone regeneration by regulating immunity,angiogenesis,and proliferation and differentiation of target cells.(3)The engineering strategies of extracellular vesicles include surface modification and cargo loading.(4)The introduction of bone morphogenetic protein 2,mutant hypoxia-inducible factor-1α,vascular endothelial growth factor,miRNA and other bioactive factors into extracellular vesicles through engineering strategies can enhance their regenerative repair ability for bone defects.

Objective:To investigate the effects of microRNA-325-3p(miR-325-3p)over-expression on the invasion and migration of colon cancer cells,and to clarify their mechanisms.Methods:The tumor tissue and the corresponding adjacent normal tissue of 25 patients clearly diagnosed with colon cancer were collected.The expression levels of miR-325-3p and relevant evolutionary and lymphoid interest domain containing protein 1(PRELID1)mRNA were detected by real-time fluorescence quantitative PCR(RT-qPCR)method.The expression levels of miR-325-3p and PRELID1 mRNA and the expression levels of PRELID1 protein in human normal colon cells NCM460 and colon cancer cells SW480,HCT116 and HT-29 were detected by RT-qPCR and Western blotting methods.The SW480 cells were divided into control group,NC-mimics group,miR-325-3p mimics group,miR-325-3p mimics+oe-NC group and miR-325-3p mimics+oe-PRELID1 group.The invasion and migration abilities of cells in various groups were detected by Transwell chamber assay and scratch healing assay,respectively.The expression levels of E-Cadherin,N-Cadherin and Vimentin in the epithelial-mesenchymal transition(EMT)of cells in various groups were detected by Western blotting method.The downstream target genes of miR-325-3p were predicted using the Targetscan bioinformatics website,and the targeted regulatory relationship between miR-325-3p and PRELID1 was verified by the dual-luciferase assay.Results:Compared with adjacent normal tissue,the expression level of miR-325-3p in cancer tissue was significantly decreased(P＜0.05),while the expression level of PRELID1 mRNA was significantly increased(P＜0.05).The expression levels of miR-325-3p and PRELID1 mRNA was negatively correlated in colon cancer tissue(r＜0,R2=0.392,P＜0.001).Compared with NCM460 cells,the expression levels of miR-325-3p in SW480,HCT116 and HT-29 cells were significantly decreased(P＜0.05),and the expression levels of PRELID1 mRNA were significantly increased(P＜0.05).Compared with control group and NC-mimics group,the number of invasive SW480 cells in miR-325-3p mimics group was significantly reduced(P＜0.05),the rate of scratch healing was decreased(P＜0.05),and the expression level of E-Cadherin protein was increased,while the expression levels of N-Cadherin and Vimentin proteins were decreased(P＜0.05).The dual-luciferase assay confirmed that PRELID1 was the direct target of miR-325-3p.Compared with miR-325-3p mimics+oe-NC group,the number of invasive SW480 cells in miR-325-3p mimics+oe-PRELID1 group was significantly increased(P＜0.05),the rate of scratch healing was increased(P＜0.05),the expression level of E-Cadherin protein was decreased,and the expression levels of N-Cadherin and Vimentin proteins were increased(P＜0.05).Conclusion:Over-expression of miR-325-3p can inhibit EMT process by down-regulating PRELID1 expression,thereby inhibiting SW480 cell invasion and migration.

The clinical antiprotozoal drug nitazoxanide has been demonstrated to improve the experimental diabetes mellitus, lipid metabolism disorders, atherosclerosis and inhibit inflammation. Since the pathogenesis of heart failure with preserved ejection (HFpEF) is multifactorial and closely associated with the aforementioned diseases, we aim to study the effect of nitazoxanide on high-fat diet (HFD) plus L-NAME (N ω-nitro-l-arginine methyl ester)-induced HFpEF and metabolic syndrome in mice. We found that oral nitazoxanide improved cardiac hypertrophy, cardiac fibrosis, cardiac diastolic dysfunction, increased blood pressure, impaired exercise tolerance, impaired glucose handling, serum lipid disorders, hepatic steatosis, increased weight of white adipose tissues and kidney fibrosis in HFD + L-NAME-treated mice. In the established HFD + L-NAME-induced HFpEF and metabolic syndrome mouse model, therapeutic treatment with nitazoxanide rescued HFD + L-NAME-induced pathological phenotypes as mentioned above. The in vitro experiments revealed that tizoxanide, the active metabolite of nitazoxanide, increased the basal mitochondria metabolism of cardiomyocytes, inhibited cardiomyocyte hypertrophy and collagen secretion from cardiac fibroblasts, and relaxed phenylephrine- and U46619-induced constriction of rat mesenteric arteries, indicating that the direct effect of tizoxanide might partly contribute to the protective effect of nitazoxanide against HFpEF in vivo. The present study suggests that nitazoxanide might be a potential drug for HFpEF and metabolic syndrome therapy.

OBJECTIVES: To study the impact of SURF4 expression level on long-term prognosis of gastric cancer (GC) and biological behaviors of GC cells. METHODS: SURF4 expression level in GC and its association with long-term patient prognosis were analyzed using publicly available databases and in 155 GC patients with low and high SURF4 expressions detected immunohistochemically. The Cox proportional hazard model and Kaplan-Meier survival curves were used to analyze independent prognostic predictors of GC and the 5-year survival rate of the patients with different SURF4 expression levels. Informatics analyses were conducted to explore the correlation of SURF4 expression level with immune cell infiltration in GC, SURF4-related differential genes and their associated pathways. In cultured GC cell line HGC-27, the effects of SURF4 knockdown and overexpression on proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) were investigated. RESULTS: Analysis of GEPIA dataset and immunohistochemical results suggested significant SURF4 overexpression in GC (P<0.05), which was associated with shortened 5-year survival time of the patients (χ²=38.749, P<0.001). The prognosis of GC was closely related to tumor stage T3-4, N2-3, CEA≥5 μg/L and CA19-9≥37 kU/L (P<0.05). SURF4 expression level was negatively correlated with activated B cells, NK cells and CD8⁺ effector memory T cells (P<0.05) and positively correlated with CD4⁺ T cells (P<0.05). GO and KEGG enrichment analysis suggested that SUFR4 may participate in GC carcinogenesis by promoting EMT through the tight junction pathway. In HGC-27 cells, SURF4 overexpression significantly decreased E-cadherin expression, increased N-cadherin expression, inhibited ZO-1 and claudin-1 expressions, and promoted cell proliferation, migration and invasion. CONCLUSIONS SURF4 is highly expressed in GC, and its overexpression is associated with a shortened 5-year survival of the patients possibly by enhancing tumor cell proliferation, migration and invasion via inhibiting tight junction proteins and promoting EMT.
Humans ; Stomach Neoplasms/metabolism* ; Cell Proliferation ; Cell Movement ; Epithelial-Mesenchymal Transition ; Cell Line, Tumor ; Neoplasm Invasiveness ; Prognosis ; Tight Junction Proteins/metabolism* ; Membrane Proteins/metabolism* ; Female ; Male

BACKGROUND:Extracellular vesicles have received extensive attention in the field of bone defect regeneration and repair in recent years.However,natural extracellular vesicles have deficiencies in sustained controlled release,tissue targeting,and drug loading capacity.Therefore,the introduction of engineering strategies to modify extracellular vesicles to enhance their therapeutic efficacy has become a research hotspot.OBJECTIVE:To review the role and application progress of engineered extracellular vesicles in the regeneration and repair of bone defects.METHODS:PubMed,Web of Science,CNKI,and WanFang databases were searched for relevant articles published in the past fifteen years.The search terms were"engineering,extracellular vesicles,exosomes,bone defect,bone regeneration,bone repair"in Chinese and English.After removal of poorly related,outdated,and duplicate studies by screening,93 articles were finally included for review according to inclusion criteria.RESULTS AND CONCLUSION:(1)Extracellular vesicles are primarily isolated based on their density,size,immunoaffinity,and surface charge.After isolation,extracellular vesicles are characterized using imaging techniques,size-and counting-based techniques,and flow cytometry.(2)Extracellular vesicles stimulate bone regeneration by regulating immunity,angiogenesis,and proliferation and differentiation of target cells.(3)The engineering strategies of extracellular vesicles include surface modification and cargo loading.(4)The introduction of bone morphogenetic protein 2,mutant hypoxia-inducible factor-1α,vascular endothelial growth factor,miRNA and other bioactive factors into extracellular vesicles through engineering strategies can enhance their regenerative repair ability for bone defects.

Objective:To investigate the current self-compassion levels in hematologic malignant tumor patients and analyze the influencing factors, providing a basis for clinical interventions.Methods:A convenience sampling method was used to select 120 patients with hematologic malignant tumors treated at the First Affiliated Hospital of Wenzhou Medical University from January to December 2022. Clinical data, self-compassion levels, social support, and adult attachment status were assessed using general data questionnaires, the Self-Compassion Scale (SCS) , the Perceived Social Support Scale (PSSS) , and the Experiences in Close Relationship Scale-Short Form (ECR-S) . Pearson correlation analysis was performed to analyze the correlation between SCS scores and PSSS, ECR-S scores. Multiple linear regression analysis was conducted to explore the influencing factors of self-compassion levels.Results:A total of 120 questionnaires were distributed, and 114 valid questionnaires were returned, resulting in an effective response rate of 95.00%. The average SCS score of hematologic malignant tumor patients was (80.54±5.80) . Univariate analysis showed that self-compassion levels differed significantly among patients of different ages, education levels, per capita family monthly income, disease duration, and the first diagnosis or not of hematologic malignant tumors ( P<0.01) . Pearson correlation analysis revealed a positive correlation between SCS scores and PSSS scores ( r=0.659, P<0.001) , and a negative correlation with the attachment anxiety and avoidance dimensions of the ECR-S ( r=-0.152, -0.588, P<0.001) . Multiple linear regression analysis showed that education level, the first diagnosis or not, PSSS score, ECR-S attachment anxiety dimension, and ECR-S attachment avoidance dimension scores entered the regression equation. Conclusions:The influencing factors of self-compassion levels in hematologic malignant tumor patients include education level, the first diagnosis or not, social support level, and adult attachment status. Healthcare providers should strengthen psychological support for patients with low education, disease recurrence/treatment failure, and insecure attachment types, while also improving patients' social support levels to enhance their self-compassion status.

Accumulating evidence has demonstrated that apoptotic vesicles(apoVs)derived from mesenchymal stem cells(MSCs;MSC-apoVs)are vital for bone regeneration,and possess superior capabilities compared to MSCs and other extracellular vesicles derived from MSCs(such as exosomes).The osteoinductive effect of MSC-apoVs is attributed to their diverse contents,especially enriched proteins or microRNAs(miRNAs).To optimize their osteoinduction activity,it is necessary to determine the unique cargo profiles of MSC-apoVs.We previously established the protein landscape and identified proteins specific to MSC-apoVs.However,the features and functions of miRNAs enriched in MSC-apoVs are unclear.In this study,we compared MSCs,MSC-apoVs,and MSC-exosomes from two types of MSC.We generated a map of miRNAs specific to MSC-apoVs and identified seven miRNAs specifically enriched in MSC-apoVs compared to MSCs and MSC-exosomes,which we classified as apoV-specific miRNAs.Among these seven specific miRNAs,hsa-miR-4485-3p was the most abundant and stable.Next,we explored its function in apoV-mediated osteoinduction.Unexpectedly,hsa-miR-4485-3p enriched in MSC-apoVs inhibited osteogenesis and promoted adipogenesis by targeting the AKT pathway.Tailored apoVs with downregulated hsa-miR-4485-3p exhibited a greater effect on bone regeneration than control apoVs.Like releasing the brake,we acquired more powerful osteoinductive apoVs.In summary,we identified the miRNA cargos,including miRNAs specific to MSC-apoVs,and generated tailored apoVs with high osteoinduction activity,which is promising in apoV-based therapies for bone regeneration.