Multimorbidity and mortality among older patients with coronary heart disease in Shenzhen,China

BACKGROUND The current understanding of the magnitude and consequences of multimorbidity in Chinese older adults with coronary heart disease(CHD)is insufficient.We aimed to assess the association and population-attributable fractions(PAFs)between multimorbidity and mortality among hospitalized older patients who were diagnosed with CHD in Shenzhen,China.METHODS We conducted a retrospective cohort study of older Chinese patients(aged≥65 years)who were diagnosed with CHD.Cox proportional hazards models were used to estimate the associations between multimorbidity and all-cause and cardiovascular disease(CVD)mortality.We also calculated the PAFs.RESULTS The study comprised 76,455 older hospitalized patients who were diagnosed with CHD between January 1,2016,and August 31,2022.Among them,70,217(91.9%)had multimorbidity,defined as the presence of at least one of the predefined 14 chronic conditions.Those with cancer,hemorrhagic stroke and chronic liver disease had the worst overall death risk,with adjusted HRs(95%CIs)of 4.05(3.77,4.38),2.22(1.94,2.53),and 1.85(1.63,2.11),respectively.For CVD mortality,the highest risk was observed for hemorrhagic stroke,ischemic stroke,and chronic kidney disease;the corresponding adjusted HRs(95%CIs)were 3.24(2.77,3.79),1.91(1.79,2.04),and 1.81(1.64,1.99),respectively.All-cause mortality was mostly attributable to cancer,heart failure and ischemic stroke,with PAFs of 11.8,10.2,and 9.1,respectively.As for CVD mortality,the leading PAFs were heart failure,ischemic stroke and diabetes;the corresponding PAFs were 18.0,15.7,and 6.1,respectively.CONCLUSIONS Multimorbidity was common and had a significant impact on mortality among older patients with CHD in Shenzhen,China.Cancer,heart failure,ischemic stroke and diabetes are the primary contributors to PAFs.Therefore,prioritizing improved treatment and management of these comorbidities is essential for the survival prognosis of CHD patients from a holistic public health perspective.

LINC_00355通过miR-15a-5p调节PHF19在肺癌侵袭转移中的作用机制研究

目的 探讨LINC_00355通过miR-15a-5p调节PHF19在肺癌侵袭转移中的作用机制。方法 采用A549肺癌细胞作为实验细胞。将细胞分为A549肺癌细胞组、lncRNA-NC组、LINC_00355 mimics组、LINC_00355 inhibitor组,进行不同处理。A549肺癌细胞组:不进行任何处理;lncRNA-NC组:加入空白载体;LINC_00355 mimics组:加入LINC_00355过表达载体;LINC_00355 inhibitor组:加入LINC_00355低表达载体。培养结束后,MTT法测定细胞活力,检测单克隆形成数、迁移和侵袭水平。采用萤光素酶报告基因实验分析LINC_00355对miR-15a-5p的靶向作用;实时荧光定量PCR(qPCR)法检测细胞LINC_00355、miR-15a-5p、PHF19,Western blot检测PHF19蛋白表达水平。结果 与A549肺癌细胞组、lncRNA-NC组比较,LINC_00355 mimics组吸光度(A)值、细胞活力、单克隆形成数、穿膜数、迁移距离升高(P<0.05),凋亡率降低(P<0.05);LINC_00355 inhibitor组A值、细胞活力、单克隆形成数、穿膜数、迁移距离降低,凋亡率升高(P<0.05);与LINC_00355 mimics组比较,LINC_00355 inhibitor组A值、细胞活力、单克隆形成数、穿膜数、迁移距离降低,凋亡率升高(P<0.05)。LINC_00355过表达可明显降低miR-15a-5p-wt的萤光素酶活性(P<0.05);与A549肺癌细胞组、lncRNA-NC组比较,LINC_00355 mimics组细胞LINC_00355、PHF19 mRNA表达上调,miR-15a-5p表达降低(P<0.05),LINC_00355 inhibitor组细胞LINC_00355及PHF19 mRNA和蛋白表达降低,miR-15a-5p表达升高(P<0.05);与LINC_00355 mimics组比较,LINC_00355 inhibitor组细胞LINC_00355 mRNA、PHF19 mRNA和蛋白表达降低,miR-15a-5p表达升高(P<0.05)。结论 LINC_00355过表达促进肺癌细胞的增殖、迁移和侵袭并诱导细胞凋亡,敲低LINC_00355则产生相反的效果;其机制可能与LINC_00355负调控miR-15a-5p进而上调肺癌细胞中PHF19的表达有关。

Construction of MnS/MoS_(2) heterostructure on two-dimensional MoS_(2) surface to regulate the reaction pathways for high-performance Li-O_(2) batteries

The inherent catalytic anisotropy of two-dimensional(2D) materials has limited the enhancement of LiO_(2) batteries(LOBs) performance due to the significantly different adsorption energies on 2D and edge surfaces.Tuning the adsorption strength in 2D materials to the reaction intermediates is essential for achieving high-performance LOBs.Herein,a MnS/MoS_(2) heterostructure is designed as a cathode catalyst by adjusting the adsorption behavior at the surface.Different from the toroidal-like discharge products on the MoS_(2) cathode,the MnS/MoS_(2) surface displays an improved adsorption energy to reaction species,thereby promoting the growth of the film-like discharge products.MnS can disturb the layer growth of MoS_(2),in which the stack edge plane features a strong interaction with the intermediates and limits the growth of the discharge products.Experimental and theoretical results confirm that the MnS/MoS_(2) heterostructure possesses improved electron transfer kinetics at the interface and plays an important role in the adsorption process for reaction species,which finally affects the morphology of Li_2O_(2),In consequence,the MnS/MoS_(2) heterostructure exhibits a high specific capacity of 11696.0 mA h g^(-1) and good cycle stability over 1800 h with a fixed specific capacity of 600 mA h g^(-1) at current density of100 mA g^(-1) This work provides a novel interfacial engineering strategy to enhance the performance of LOBs by tuning the adsorption properties of 2D materials.

Fine mapping and cloning of the sterility gene Bra2Ms in nonheading Chinese cabbage(Brassica rapa ssp.chinensis)

The application of a male-sterile line is an ideal approach for hybrid seed production in non-heading Chinese cabbage(Brassica rapa ssp.chinensis).However,the molecular mechanisms underlying male sterility in B.rapa are still largely unclear.We previously obtained the natural male sterile line WS24-3 of non-heading Chinese cabbage and located the male sterile locus,Bra2Ms,on the A2 chromosome.Cytological observations revealed that the male sterility of WS24-3 resulted from disruption of the meiosis process during pollen formation.Fine mapping of Bra2Ms delimited the locus within a physical distance of about 129 kb on the A2 chromosome of B.rapa.The Bra039753 gene encodes a plant homeodomain(PHD)-finger protein and is considered a potential candidate gene for Bra2Ms.Bra039753 was significantly downregulated in sterile line WS24-3 compared to the fertile line at the meiotic anther stage.Sequence analysis of Bra039753 identified a 369 bp fragment insertion in the first exon in male sterile plants,which led to an amino acid insertion in the Bra039753 protein.In addition,the 369 bp fragment insertion was found to cosegregate with the male sterility trait.This study identified a novel locus related to male sterility in non-heading Chinese cabbage,and the molecular marker obtained in this study will be beneficial for the marker-assisted selection of excellent sterile lines in non-heading Chinese cabbage and other Brassica crops.

Improved viability of trehalose on Lactobacillus plantarum embedded with whey protein concentrate/pullulan in simulated gastrointestinal conditions and its application in acid juice

Trehalose(TRE)was used to improve the gastrointestinal tolerance of Lactobacillus plantarum embedded with whey protein concentrate/pullulan(WPC/PUL)hydrogel and the embedded L.plantarum was applied to juice.The study indicated that 5%TRE significantly increased the viable counts of L.plantarum embedded in WPC/PUL hydrogel from(8.83±0.03)to(9.14±0.04)(lg(CFU/g))in simulated gastric juice(SGJ)and from(9.13±0.04)to(9.38±0.04)(lg(CFU/g))in simulated intestinal juice,respectively.The addition of TRE improved the glass transition temperature of WPC/PUL hydrogel and decreased the hardness and its solubility in SGJ,which may be responsible for the improved protection of WPC/PUL hydrogels on L.plantarum.In addition,TRE increased the viable counts of L.plantarum in WPC/PUL probiotic microcapsule juice at low pH and high temperature during storage.

Based on non-targeted metabolomics for differential components screening of Rosae Chinensis Flos and Rosae Rugosae Flos and their quality evaluation

Background:Rosa chinensis Jacq.and Rosa rugosa Thunb.are not only of ornamental value,but also edible flowers and the flower buds have been listed in the Chinese Pharmacopoeia as traditional medicines.The two plants have some differences in efficacy,but the flower buds are easily confused for similar traits.In addition,large-scale cultivation of ornamental rose flowers may lead to a decrease in the effective components of medicinal roses.Therefore,it is necessary to study the chemical composition and make quality evaluation of Rosae Chinensis Flos(Yueji)and Rosae Rugosae Flos(Meigui).Methods:In this study,40 batches of samples including Meigui and Yueji from different regions in China were collected to establish high-performance liquid chromatography fingerprints.Then,the fingerprints data was analyzed using principal component analysis,hierarchical cluster analysis,and partial least squares discriminant analysis analysis chemometrics to obtain information on intergroup differences,and non-targeted metabolomic techniques were applied to identify and compare chemical compositions of samples which were chosen from groups with large differences.Differential compounds were screened by orthogonal partial least-squares discriminant analysis and S-plot,and finally multi-component quantification was performed to comprehensively evaluate the quality of Yueji and Meigui.Results:The similarity between the fingerprints of 40 batches roses and the reference print R was 0.73 to 0.93,indicating that there were similarities and differences between the samples.Through principal component analysis and hierarchical cluster analysis of fingerprints data,the samples from different origins and varieties were intuitively divided into four groups.Partial least-squares discriminant analysis analysis showed that Meigui and Yueji cluster into two categories and the model was reliable.A total of 89 compounds were identified by high resolution mass spectrometry,mainly were flavonoids and flavonoid glycosides,as well as phenolic acids.Eight differential components were screened out by orthogonal partial least-squares discriminant analysis and S-plot analysis.Quantitative analyses of the eight compounds,including gallic acid,ellagic acid,hyperoside,isoquercitrin,etc.,showed that Yueji was generally richer in phenolic acids and flavonoids than Meigui,and the quality of Yueji from Shandong and Hebei was better.It is worth noting that Xinjiang rose is rich in various components,which is worth focusing on more in-depth research.Conclusion:In this study,the fingerprints of Meigui and Yueji were established.The chemical components information of roses was further improved based on non-targeted metabolomics and mass spectrometry technology.At the same time,eight differential components of Meigui and Yueji were screened out and quantitatively analyzed.The research results provided a scientific basis for the quality control and rational development and utilization of Rosae Chinensis Flos and Rosae Rugosae Flos,and also laid a foundation for the study of their pharmacodynamic material basis.

Transcriptomic Analysis Reveals the Formation Mechanism of Anthocyanins Light-Independent Synthesis in Chrysanthemum

Chrysanthemum×morifolium is a horticultural crop which plays a vital role in theflower industry with signifi-cant economic value and has a cultivation history of over three thousand years in China.The accumulation of anthocyanins is always affected by light.Here,we revealed that anthocyanin accumulation is highly dependent on light in‘2021135’genotype chrysanthemum,while it is light-independent in‘2001402’genotype chrysanthe-mum.However,no literature has been reported regarding the non-photosensitive chrysanthemum in anthocya-nins light-independent synthesis pathways.Through the phenotype analysis of 44 F1 generations,we found that light-independence is a dominant trait which can be stable inherited by progeny.The transcriptome of the rayflorets of‘2021135’and‘2001402’under light and bagging treatment were sequenced and analyzed.Based on weighted gene co-expression network analysis(WGCNA),K-means analysis,and Real-Time Quantitative Poly-merase Chain Reaction(RT-qPCR)analysis,16 genes were highly correlated with the anthocyanin content.The anthocyanin content of rayflorets treated with different light-quality conditions indicated that blue light signifi-cantly affected anthocyanin accumulations.Through Yeast one-hybrid analysis,CmBIC1.1 and CmBIC1.2 can directly regulate the anthocyanin structural gene CmCHS2.In our study,we revealed the important characteristics of light-independent anthocyanin synthesis in chrysanthemums and screened regulatory factors in light-depen-dent and light-independent anthocyanin synthesis pathways.The results laid the groundwork for subsequent ana-lysis of the molecular mechanism involved in the light-independent synthesis of anthocyanins in chrysanthemums.

Metformin administration in prevention of colorectal polyps in type 2 diabetes mellitus patients

BACKGROUND Colorectal polyps are frequently observed in patients with type 2 diabetes mellitus(DM),posing a significant risk for colorectal cancer.Metformin,a widely prescribed biguanidine drug for type 2 DM,has been suggested to have potential chemoprophylactic effects against various cancers.AIM To explore the correlation between colorectal polyps and metformin use in type 2 DM patients.METHODS Type 2 DM patients were categorized into polyp and non-polyp groups.Following this,all patients were categorized into the type 2 DM-metformin,type 2 DM-non-metformin,and non-type 2 DM groups.Based on the baseline colonoscopy results,we performed pairwise comparisons of the incidence of colorectal polyps among the three groups.Additionally,we analyzed the relationship between colorectal polyps and the duration of metformin use and between the size and number of polyps and metformin use.Simultaneously,we focused on the specific pathological types of polyps and analyzed their relationship with metformin use.Finally,we compared the incidence of polyps between metformin and non-metformin groups according to the interval colonoscopy results.RESULTS The rate of metformin use in patients with colorectal polyps was 0.502 times that of patients without colorectal polyps[odds ratio(OR)=0.502,95%confidence interval(CI):0.365-0.689;P<0.001].The incidence of colorectal polyps did not differ significantly between the type 2 DM-metformin and non-type 2 DM groups(P>0.05).Furthermore,the correlations between the duration of metformin use and the incidence of colorectal polyps and between the size and number of polyps and metformin use were not statistically significant(P>0.05).Metformin use did not affect the incidence of colorectal polyps during interval colonoscopy(P>0.05).CONCLUSION Metformin use and colorectal polyp incidence in type 2 DM patients showed a negative correlation,independent of the hypoglycemic effect of metformin.

The cardiovascular system at high altitude:A bibliometric and visualization analysis

BACKGROUND When exposed to high-altitude environments,the cardiovascular system undergoes various changes,the performance and mechanisms of which remain controversial.AIM To summarize the latest research advancements and hot research points in the cardiovascular system at high altitude by conducting a bibliometric and visualization analysis.METHODS The literature was systematically retrieved and filtered using the Web of Science Core Collection of Science Citation Index Expanded.A visualization analysis of the identified publications was conducted employing CiteSpace and VOSviewer.RESULTS A total of 1674 publications were included in the study,with an observed annual increase in the number of publications spanning from 1990 to 2022.The United States of America emerged as the predominant contributor,while Universidad Peruana Cayetano Heredia stood out as the institution with the highest publication output.Notably,Jean-Paul Richalet demonstrated the highest productivity among researchers focusing on the cardiovascular system at high altitude.Furthermore,Peter Bärtsch emerged as the author with the highest number of cited articles.Keyword analysis identified hypoxia,exercise,acclimatization,acute and chronic mountain sickness,pulmonary hypertension,metabolism,and echocardiography as the primary research hot research points and emerging directions in the study of the cardiovascular system at high altitude.CONCLUSION Over the past 32 years,research on the cardiovascular system in high-altitude regions has been steadily increasing.Future research in this field may focus on areas such as hypoxia adaptation,metabolism,and cardiopulmonary exercise.Strengthening interdisciplinary and multi-team collaborations will facilitate further exploration of the pathophysiological mechanisms underlying cardiovascular changes in high-altitude environments and provide a theoretical basis for standardized disease diagnosis and treatment.

Additive manufacturing of promising heterostructure for biomedical applications

As a new generation of materials/structures,heterostructure is characterized by heterogeneous zones with dramatically different mechanical,physical or chemical properties.This endows heterostructure with unique interfaces,robust architectures,and synergistic effects,making it a promising option as advanced biomaterials for the highly variable anatomy and complex functionalities of individual patients.However,the main challenges of developing heterostructure lie in the control of crystal/phase evolution and the distribution/fraction of components and structures.In recent years,additive manufacturing techniques have attracted increasing attention in developing heterostructure due to the unique flexibility in tailored structures and synthetic multimaterials.This review focuses on the additive manufacturing of heterostructure for biomedical applications.The structural features and functional mechanisms of heterostructure are summarized.The typical material systems of heterostructure,mainly including metals,polymers,ceramics,and their composites,are presented.And the resulting synergistic effects on multiple properties are also systematically discussed in terms of mechanical,biocompatible,biodegradable,antibacterial,biosensitive and magnetostrictive properties.Next,this work outlines the research progress of additive manufacturing employed in developing heterostructure from the aspects of advantages,processes,properties,and applications.This review also highlights the prospective utilization of heterostructure in biomedical fields,with particular attention to bioscaffolds,vasculatures,biosensors and biodetections.Finally,future research directions and breakthroughs of heterostructure are prospected with focus on their more prospective applications in infection prevention and drug delivery.

18β-glycyrrhetinic acid inhibits proliferation of gastric cancer cells through regulating the miR-345-5p/TGM2 signaling pathway

BACKGROUND Gastric cancer(GC)is a common gastrointestinal malignancy worldwide.Based on cancer-related mortality,the current prevention and treatment strategies for GC still show poor clinical results.Therefore,it is important to find effective drug treatment targets.AIM To explore the molecular mechanism of 18β-glycyrrhetinic acid(18β-GRA)regulating the miR-345-5p/TGM2 signaling pathway to inhibit the proliferation of GC cells.METHODS CCK-8 assay was used to determine the effect of 18β-GRA on the survival rate of GES-1 cells and AGS and HGC-27 cells.Cell cycle and apoptosis were detected by flow cytometry,cell migration was detected by a wound healing assay,the effect of 18β-GRA on subcutaneous tumor growth in BALB/c nude mice was investigated,and the cell autophagy level was determined by MDC staining.TMT proteomic analysis was used to detect the differentially expressed autophagy-related proteins in GC cells after 18β-GRA intervention,and then the protein-protein interaction was predicted using STRING(https://string-db.org/).MicroRNAs(miRNAs)transcriptome analysis was used to detect the miRNA differential expression profile,and use miRBase(https://www.mirbase/)and TargetScan(https://www.targetscan.org/)to predict the miRNA and complementary binding sites.Quantitative real-time polymerase chain reaction was used to detect the expression level of miRNA in 18β-GRA treated cells,and western blot was used to detect the expression of autophagy related proteins.Finally,the effect of miR-345-5p on GC cells was verified by mir-345-5p overexpression.RESULTS 18β-GRA could inhibit GC cells viability,promote cell apoptosis,block cell cycle,reduce cell wound healing ability,and inhibit the GC cells growth in vivo.MDC staining results showed that 18β-GRA could promote autophagy in GC cells.By TMT proteomic analysis and miRNAs transcriptome analysis,it was concluded that 18β-GRA could down-regulate TGM2 expression and up-regulate miR-345-5p expression in GC cells.Subsequently,we verified that TGM2 is the target of miR-345-5p,and that overexpression of miR-345-5p significantly inhibited the protein expression level of TGM2.Western blot showed that the expression of autophagy-related proteins of TGM2 and p62 was significantly reduced,and LC3II,ULK1 and AMPK expression was significantly increased in GC cells treated with 18β-GRA.Overexpression of miR-345-5p not only inhibited the expression of TGM2,but also inhibited the proliferation of GC cells by promoting cell apoptosis and arresting cell cycle.CONCLUSION 18β-GRA inhibits the proliferation of GC cells and promotes autophagy by regulating the miR-345-5p/TGM2 signaling pathway.

MiR-204-3p overexpression inhibits gastric carcinoma cell proliferation by inhibiting the MAPK pathway and RIP1/MLK1 necroptosis pathway to promote apoptosis

BACKGROUND Gastric carcinoma(GC)is the third most frequent cause of cancer-related death,highlighting the pressing need for novel clinical treatment options.In this regard,microRNAs(miRNAs)have emerged as a promising therapeutic strategy.Studies have shown that miRNAs can regulate related signaling pathways,acting as tumor suppressors or tumor promoters.AIM To explore the effect of miR-204-3p on GC cells.METHODS We measured the expression levels of miR-204-3p in GC cells using quantitative real-time polymerase chain reaction,followed by the delivery of miR-204-3p overexpression and miR-204-3p knockdown vectors into GC cells.CCK-8 was used to detect the effect of miR-204-3p on the proliferation of GC cells,and the colony formation ability of GC cells was detected by the clonal formation assay.The effects of miR-204-3p on GC cell cycle and apoptosis were detected by flow cytometry.The BABL/c nude mouse subcutaneous tumor model using MKN-45 cells was constructed to verify the effect of miR-204-3p on the tumorigenicity of GC cells.Furthermore,the study investigated the effects of miR-204-3p on various proteins related to the MAPK signaling pathway,necroptosis signaling pathway and apoptosis signaling pathway on GC cells using Western blot techniques.RESULTS Firstly,we found that the expression of miR-204-3p in GC was low.When treated with the lentivirus overexpression vector,miR-204-3p expression significantly increased,but the lentivirus knockout vector had no significant effect on miR-204-3p.In vitro experiments confirmed that miR-204-3p overexpression inhibited GC cell viability,promoted cell apoptosis,blocked the cell cycle,and inhibited colony formation ability.In vivo animal experiments confirmed that miR-204-3p overexpression inhibited subcutaneous tumorigenesis ability in BABL/c nude mice.Simultaneously,our results verified that miR-204-3p overexpression can inhibit GC cell proliferation by inhibiting protein expression levels of KRAS and p-ERK1/2 in the MAPK pathway,as well as inhibiting protein expression levels of p-RIP1 and p-MLK1 in the necroptosis pathway to promote the BCL-2/BAX/Caspase-3 apoptosis pathway.CONCLUSION MiR-204-3p overexpression inhibited GC cell proliferation by inhibiting the MAPK pathway and necroptosis pathway to promote apoptosis of GC cells.Thus,miR-204-3p may represent a new potential therapeutic target for GC.

先天性常染色体隐性遗传性鱼鳞病

目的:分析1例TGM1基因变异引起的先天性常染色体隐性遗传性鱼鳞病(ARCI)临床表现和基因型。方法:对患儿进行TGM1基因测序。应用Sanger测序法对可疑致病位点进行家系验证。结果:羊水检测示TGM1基因c.968G>A和c.871G>A复合杂合变异,分别遗传自无表型的父亲及母亲。患儿出生时皮肤潮红呈火棉胶样改变,经保湿和润肤等治疗皮损改善。结论:TGM1基因c.968G>A和c.871G>A复合杂合变异可能是患儿的致病原因。

拔罐联合瘦腹泄浊方火疗对痰湿型肥胖症患者的效果观察

目的 观察拔罐联合瘦腹泄浊方火疗对痰湿型肥胖症患者的临床疗效。方法 选取2022年7月至2023年7月在江苏省某中医院内分泌科进行治疗的痰湿型肥胖症患者60例为研究对象,根据入院时间先后分为对照组和观察组,每组各30例。对照组予生活方式干预,包括饮食、运动及健康教育干预,观察组在对照组干预的基础上采用拔罐联合瘦腹泄浊方火疗,1个月为1个疗程,共治疗2个疗程。比较两组患者干预前后肥胖相关指标、中医证候积分及疗效。结果 两组患者治疗后体重、腰围、体质指数、中医症候积分等均降低,且观察组低于对照组,差异有统计学意义(P<0.05)。结论 拔罐联合瘦腹泄浊方火疗可以减少痰湿型肥胖症患者体重、腰围,降低其体质指数、体脂率,提高患者的生活质量。

18β-glycyrrhetinic acid promotes gastric cancer cell autophagy and inhibits proliferation by regulating miR-328-3p/signal transducer and activator of transcription 3

BACKGROUND Gastric cancer(GC)is one of the most common cancer types worldwide,and its prevention and treatment methods have garnered much attention.As the active ingredient of licorice,18β-glycyrrhetinic acid(18β-GRA)has a variety of pharmacological effects.The aim of this study was to explore the effective target of 18β-GRA in the treatment of GC,in order to provide effective ideas for the clinical prevention and treatment of GC.AIM To investigate the mechanism of 18β-GRA in inhibiting cell proliferation and promoting autophagy flux in GC cells.METHODS Whole transcriptomic analyses were used to analyze and screen differentially expressed microRNAs(miRNAs)in GC cells after 18β-GRA intervention.Lentivirus-transfected GC cells and the Cell Counting Kit-8 were used to detect cell proliferation ability,cell colony formation ability was detected by the clone formation assay,and flow cytometry was used to detect the cell cycle and apoptosis.A nude mouse transplantation tumor model of GC cells was constructed to verify the effect of miR-328-3p overexpression on the tumorigenicity of GC cells.Tumor tissue morphology was observed by hematoxylin and eosin staining,and microtubule-associated protein light chain 3(LC3)expression was detected by immunohistochemistry.TransmiR,STRING,and miRWalk databases were used to predict the relationship between miR-328-3p and signal transducer and activator of transcription 3(STAT3)-related information.Expression of STAT3 mRNA and miR-328-3p was detected by quantitative polymerase chain reaction(qPCR)and the expression levels of STAT3,phosphorylated STAT3(p-STAT3),and LC3 were detected by western blot analysis.The targeted relationship between miR-328-3p and STAT3 was detected using the dual-luciferase reporter gene system.AGS cells were infected with monomeric red fluorescent protein-green fluorescent protein-LC3 adenovirus double label.LC3 was labeled and autophagy flow was observed under a confocal laser microscope.RESULTS The expression of miR-328-3p was significantly upregulated after 18β-GRA intervention in AGS cells(P=4.51E-06).Overexpression of miR-328-3p inhibited GC cell proliferation and colony formation ability,arrested the cell cycle in the G0/G1 phase,promoted cell apoptosis,and inhibited the growth of subcutaneous tumors in BALB/c nude mice(P<0.01).No obvious necrosis was observed in the tumor tissue in the negative control group(no drug intervention or lentivirus transfection)and vector group(the blank vector for lentivirus transfection),and more cells were loose and necrotic in the miR-328-3p group.Bioinformatics tools predicted that miR-328-3p has a targeting relationship with STAT3,and STAT3 was closely related to autophagy markers such as p62.After overexpressing miR-328-3p,the expression level of STAT3 mRNA was significantly decreased(P<0.01)and p-STAT3 was downregulated(P<0.05).The dual-luciferase reporter gene assay showed that the luciferase activity of miR-328-3p and STAT33’untranslated regions of the wild-type reporter vector group was significantly decreased(P<0.001).Overexpressed miR-328-3p combined with bafilomycin A1(Baf A1)was used to detect the expression of LC3 II.Compared with the vector group,the expression level of LC3 II in the overexpressed miR-328-3p group was downregulated(P<0.05),and compared with the Baf A1 group,the expression level of LC3 II in the overexpressed miR-328-3p+Baf A1 group was upregulated(P<0.01).The expression of LC3 II was detected after intervention of 18β-GRA in GC cells,and the results were consistent with the results of miR-328-3p overexpression(P<0.05).Additional studies showed that 18β-GRA promoted autophagy flow by promoting autophagosome synthesis(P<0.001).qPCR showed that the expression of STAT3 mRNA was downregulated after drug intervention(P<0.05).Western blot analysis showed that the expression levels of STAT3 and p-STAT3 were significantly downregulated after drug intervention(P<0.05).CONCLUSION 18β-GRA promotes the synthesis of autophagosomes and inhibits GC cell proliferation by regulating the miR-328-3p/STAT3 signaling pathway.

Data analysis guidelines for single‑cell RNA‑seq in biomedical studies and clinical applications

The application of single-cell RNA sequencing(scRNA-seq)in biomedical research has advanced our understanding of the pathogenesis of disease and provided valuable insights into new diagnostic and therapeutic strategies.With the expansion of capacity for high-throughput scRNA-seq,including clinical samples,the analysis of these huge volumes of data has become a daunting prospect for researchers entering this field.Here,we review the workflow for typical scRNA-seq data analysis,covering raw data processing and quality control,basic data analysis applicable for almost all scRNA-seq data sets,and advanced data analysis that should be tailored to specific scientific questions.While summarizing the current methods for each analysis step,we also provide an online repository of software and wrapped-up scripts to support the implementation.Recommendations and caveats are pointed out for some specific analysis tasks and approaches.We hope this resource will be helpful to researchers engaging with scRNA-seq,in particular for emerging clinical applications.

Generalization of PhaseNet in Shandong and its application to the Changqing M4.1 earthquake sequence

Waveforms of seismic events,extracted from January 2019 to December 2021 were used to construct a test dataset to investigate the generalizability of PhaseNet in the Shandong region.The results show that errors in the picking of seismic phases(P-and Swaves)had a broadly normal distribution,mainly concentrated in the ranges of−0.4–0.3 s and−0.4–0.8 s,respectively.These results were compared with those published in the original PhaseNet article and were found to be approximately 0.2–0.4 s larger.PhaseNet had a strong generalizability for P-and S-wave picking for epicentral distances of less than 120 km and 110 km,respectively.However,the phase recall rate decreased rapidly when these distances were exceeded.Furthermore,the generalizability of PhaseNet was essentially unaffected by magnitude.The M4.1 earthquake sequence in Changqing,Shandong province,China,that occurred on February 18,2020,was adopted as a case study.PhaseNet detected more than twice the number of earthquakes in the manually obtained catalog.This further verified that PhaseNet has strong generalizability in the Shandong region,and a high-precision earthquake catalog was constructed.According to these precise positioning results,two earthquake sequences occurred in the study area,and the southern cluster may have been triggered by the northern cluster.The focal mechanism solution,regional stress field,and the location results of the northern earthquake sequence indicated that the seismic force of the earthquake was consistent with the regional stress field.

Comparison of polysaccharides from 10 species of genera Paris,Trillium,Aspidistra,and Polygonatum

Background:The genera Paris,Trillium,Aspidistra,and Polygonatum originally belonged to the family Liliaceae;however,recently,in the APG system,the genera Polygonatum and Aspidistra were placed in the family Asparagaceae and the genera Paris and Trillium were placed in the family Melanthiaceae.Methods:To study the application of polysaccharide chemotaxonomy in Asparagaceae and Melanthiaceae,we extracted polysaccharides from the rhizomes of plants,including Paris polyphylla var.yunnanensis,Paris daliensis,Paris polyphylla var.chinensis,Trillium tschonoskii,Aspidistra elatior,Aspidistra sichuanensis,Aspidistra caespitosa,Polygonatum sibiricum,Polygonatum cyrtonema,and Polygonatum kingianum.Physicochemical and structural characterization of these polysaccharides was performed using molecular weight,monosaccharide composition,Fourier-transform infrared spectroscopy,nuclear magnetic resonance spectroscopy,and scanning electron microscopy.The immunomodulatory activities were evaluated using the macrophage cell line,RAW264.7.Results:In this study,we found that the polysaccharides from the genus Polygonatum and Aspidistra were mainly comprised of fructose and glucose;the molecular weights were mostly concentrated between 4.6-6.2 kDa.The polysaccharides from species of genus Polygonatum differed in that they contained acetyl groups,while polysaccharides from species of genus Aspidistra did not.Paris and Trillium species polysaccharides mainly consisted of glucose and mannose;the molecular weights of the two major peaks were 6.3-13 kDa and 1-1.5 kDa,respectively;the main differences between polysaccharides were the ratio of glucose and mannose and the degree of branching.These results suggest that fructans might be the main feature of the family Asparagaceae and fructans with acetyl groups might be the main feature of the genus Polygonatum which is consistent with the results of previous studies.Additionally,all polysaccharides from the 10 plants promoted phagocytic activity.Some of them exhibited strong activity,including nitrogen monoxide release and TNF-αsecretion by RAW264.7 macrophages,indicating that they could be used as potential immunomodulatory agents.Conclusion:The molecular weight distribution,monosaccharide composition,and surface morphology of the polysaccharides of the Polygonatum and Aspidistra plants were similar,while those of Paris and Trillium plants were similar.These results support the APG classification system,which placed the genera Polygonatum and Aspidistra in the Asparagaceae family and placed the genera Paris and Trillium in the Melanthiaceae family.

A review on the extraction and separation of andrographolide from Andrographis paniculata:extraction selectivity,current challenges and strategies

Andrographolide is the main active ingredient of Andrographis paniculata(Burm.f.)Nees,known as“natural antibiotic”.Here,for the purpose of discovering a more efficient,low-cost extraction and separation method,the research status of andrographolide was reviewed.At present,researches only take extraction rate as the only index to optimize extraction parameters,but ignores the importance of extraction selectivity.It is usually meaningless to blindly pursue the extraction rate without considering the difficulty and cost of subsequent separation.So,factors affecting extraction selectivity such as solvent choice,temperature and physicochemical effects caused by extraction technique itself,are first discussed.Different extraction techniques for andrographolide were discussed by comparing the selectivity,efficiency and cost of extraction.The separation procedures of andrographolide such as decolorization,impurity removal,crystallization,membrane separation,solid-phase extraction and partition chromatography and their challenges and possible strategies are also discussed.It is hoped that this review can provide guidance for researchers who are committed to advancing the field of andrographolide extraction and purification.

Effect of salvianolic acid B-loaded mesoporous silica nanoparticles on myocardial ischemia-reperfusion injury

Background:Currently,no drugs can specifically improve clinical cardiac ischemia-reperfusion injury or the prognosis of hemodialysis.Salvianolic acid B(SalB)is a widely used cardiac protectant;however,its clinical application is limited by its low oral bioavailability and poor intestinal absorption.The exploration of its preparation and clinical applications has become a research hotspot in recent years.Methods:To determine whether mesoporous silica nanoparticles(MSNs)efficiently delivered SalB to the heart and SalB@MSNs-RhB reduced myocardial ischemia-reperfusion injury,we constructed a myocardial ischemia-reperfusion male rat model,hypoxia/reoxygenation cardiomyocytes,and treated them with SalB@MSNs-RhB.Results:SalB@MSNs-RhB showed improved bioavailability,therapeutic effect,heightened JAK2/STAT3-dependent pro-survival signaling,and antioxidant responses,thereby protecting cardiomyocytes from ischemia-reperfusion injury-induced oxidative stress and apoptosis.Conclusion:This use of SalB-loaded nanoparticles and investigation of their mechanism of action may provide a new strategy for treating cardiomyocytes.Thus,hypoxia/reoxygenation promotes the clinical application of SalB.