Gut microbiota contributes to the distinction between two traditional Chinese medicine syndromes of ulcerative colitis

BACKGROUND Ulcerative colitis(UC)is considered to be closely associated with alteration of intestinal microorganisms.According to the traditional Chinese medicine(TCM)theory,UC can be divided into two disease syndromes called Pi-Xu-Shi-Yun(PXSY)and Da-Chang-Shi-Re(DCSR).The relationships among gut microbiota,TCM syndromes,and UC pathogenesis have not been well investigated.AIM To investigate the role of gut microbiota in UC and the distinction of microbiota dysbiosis between PXSY and DCSR syndromes.METHODS From May 2015 to February 2016,UC patients presenting to LongHua Hospital who met the established inclusion and exclusion criteria were enrolled in this retrospective study.Fresh stool specimens of UC patients with PXSY or DCSR were collected.The feces of the control group came from the health examination population of Longhua Hospital.The composition of gut bacterial communities in stool samples was determined by the pyrosequencing of 16S ribosomal RNA.The high-throughput sequencing reads were processed with QIIME,and biological functions were predicted using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States.RESULTS The composition of gut bacterial communities in 93 stool samples(30 healthy controls,32 patients with PXSY syndrome,and 31 patients with DCSR syndrome)was determined by the pyrosequencing of 16S ribosomal RNA.Beta diversity showed that the composition of the microbiota was different among the three groups.At the family level,Porphyromonadaceae,Rikeneliaceae,and Lachnospiraceae significantly decreased while Enterococcus,Streptococcus,and other potential pathogens significantly increased in UC patients compared to healthy subjects.At the genus level,Parabacteroides,Dorea,and Ruminococcus decreased while Faeca-libacterium showed increased abundance in UC compared to healthy controls.Five differential taxa were identified between PXSY and DCSR syndromes.At the genus level,a significantly increased abundance of Streptococcus was observed in DCSR patients,while Lachnoclostridium increased in PXSY patients.The differential functional pathways of the gut microbiome between the PXSY and DCSR groups mainly included lipid metabolism,immunity,and the metabolism of polypeptides.CONCLUSION Our study suggests that the gut microbiota contributes to the distinction between the two TCM syndromes of UC.

Jianpi Qingchang decoction alleviates ulcerative colitis by inhibiting nuclear factor-κB activation

To investigate the therapeutic effect of Jianpi Qingchang decoction (JPQCD) on dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice.METHODSC57BL/c mice were injected intragastrically with 5% DSS instead of drinking water for 7 d, and their body weight, diarrhea severity and fecal bleeding were monitored, while the mice in the control group were treated with standard drinking water, without DSS. After 7 d, the DSS drinking water was changed to normal water and the DSS group continued with DSS water. The control and DSS groups were given normal saline by intragastric injection. The 5-aminosalicylic acid (5-ASA) group was treated orally with 5-ASA at a dose of 100 mg/kg daily. The JPQCD group was treated orally with JPQCD at a dose of 17.1 g/kg daily. On day 14, the colon length was measured, the colorectal histopathological damage score was assessed, and protein levels of interleukin (IL)-1β, IL-8 and tumor necrosis factor-alpha (TNF-α) in colon supernatants were measured by enzyme-linked immunosorbent assay. mRNA expression of IL-1β, IL-8, TNF-α and nuclear factor-kappa B (NF-κB) was detected by real-time quantitative polymerase chain reaction. Western blotting was used to detect the protein expression of NF-κB and inhibitor of kappa B.RESULTSAcute inflammation occurred in the mice administered DSS, including the symptoms of losing body weight, loose feces/watery diarrhea and presence of fecal blood; all these symptoms worsened at 7 d. The colons of mice treated with DSS were assessed by histological examination, and the results confirmed that acute inflammation had occurred, as evidenced by loss of colonic mucosa and chronic inflammatory cell infiltration, and these features extended into the deeper layer of the colon walls. The expression levels of IL-1β, IL-8 and TNF-α in the DSS group were higher than those in the control group (P < 0.05), and the expression levels of IL-1β, IL-8 and TNF-α in the JPQCD and 5-ASA groups were lower than those in the DSS group after treating with JPQCD and 5-ASA. Comparing with the DSS group, the mRNA level of IL-1β, IL-8, TNF-α and NF-κB was significantly reduced by 5-ASA and JPQCD. The difference between JPQCD and 5-ASA groups was not statistically significant (P > 0.05). Comparing with the DSS group, due to using JPQCD and 5-ASA, significant suppression of activation in DSS-induced NF-κB and increased phosphorylation of IκB in mice with experimental colitis occurred (P < 0.05). The difference between the JPQCD group and the 5-ASA group was not statistically significant (P > 0.05).CONCLUSIONActivation of the NF-κB signaling pathway is inhibited by JPQCD, which shows the potential mechanism by which JPQCD treats UC.

Jianpi Qingchang decoction regulates intestinal motility of dextran sulfate sodium-induced colitis through reducing autophagy of interstitial cells of Cajal

AIM To investigate the underlying effect of Jianpi Qingchang decoction(JQD) regulating intestinal motility of dextran sulfate sodium(DSS)-induced colitis in mice. METHODS C57BL/6 mice were randomly divided into four groups: the control group, the DSS group, the JQD group, and the 5-aminosalicylic acid group. Except for the control group, colitis was induced in other groups by giving distilled water containing 5% DSS. Seven days after modeling, the mice were administered corresponding drugs intragastrically. The mice were sacrificed on the 15^(th) day. The disease activity index, macroscopic and histopathologic lesions, and ultrastructure of colon interstitial cells of Cajal(ICC) were observed. The levels of tumor necrosis factor-alpha(TNF-α), interleukin(IL)-1β, IL-10 and interferon gamma(IFN-γ), the expression of nuclear factor-kappa B(NF-κB) p65, c-kit, microtubule-associated protein 1 light chain 3(LC3-Ⅱ) and Beclin-l m RNA, and the colonic smooth muscle tension were assessed. RESULTS Acute inflammation occurred in the mice administered DSS. Compared with the control group, the levels of IL-1β, TNF-α, IL-10 and IFN-γ, the expression of LC3-Ⅱ, Beclin-1 and NF-κB p65 m RNA, and the contractile frequency increased(P < 0.05), the expression of c-kit m RNA and the colonic smooth muscle contractile amplitude decreased in the DSS group(P < 0.05). Compared with the DSS group, the levels of IL-10 and IFN-γ, the expression of c-kit m RNA, and the colonic smooth muscle contractile amplitude increased(P < 0.05), the levels of TNF-α and IL-1β, the expression of LC3-Ⅱ, Beclin-1 and NF-κB p65 m RNA, and the contractile frequency decreased in the JQD group(P < 0.05).CONCLUSION JQD can regulate the intestinal motility of DSS-induced colitis in mice through suppressing intestinal inflammatory cascade reaction, reducing autophagy of ICC, and regulating the network path of ICC/smooth muscle cells.

Fecal microbiota transplantation in the metabolic diseases:Current status and perspectives

With the development of microbiology and metabolomics,the relationship between the intestinal microbiome and intestinal diseases has been revealed.Fecal microbiota transplantation(FMT),as a new treatment method,can affect the course of many chronic diseases such as metabolic syndrome,malignant tumor,autoimmune disease and nervous system disease.Although the mechanism of action of FMT is now well understood,there is some controversy in metabolic diseases,so its clinical application may be limited.Microflora transplantation is recommended by clinical medical guidelines and consensus for the treatment of recurrent or refractory Clostridium difficile infection,and has been gradually promoted for the treatment of other intestinal and extraintestinal diseases.However,the initial results are varied,suggesting that the heterogeneity of the donor stools may affect the efficacy of FMT.The success of FMT depends on the microbial diversity and composition of donor feces.Therefore,clinical trials may fail due to the selection of ineffective donors,and not to faulty indication selection for FMT.A new understanding is that FMT not only improves insulin sensitivity,but may also alter the natural course of type 1 diabetes by modulating autoimmunity.In this review,we focus on the main mechanisms and deficiencies of FMT,and explore the optimal design of FMT research,especially in the field of cardiometabolic diseases.

Gut microbiota and inflammatory bowel disease: The current status and perspectives

Inflammatory bowel disease(IBD)is a chronic immune-mediated disease that affects the gastrointestinal tract.It is argued that environment,microbiome,and immune-mediated factors interact in a genetically susceptible host to trigger IBD.Recently,there has been increased interest in the development,progression,and treatment of IBD because of our understanding of the microbiome.Researchers have proved that some factors can alter the microbiome and the pathogenesis of IBD.As a result,there has been increasing interest in the application of probiotics,prebiotics,antibiotics,fecal microbiota transplantation,and gene manipulation in treating IBD because of the possible curative effect of microbiome-modulating interventions.In this review,we summarize the findings from human and animal studies and discuss the effect of the gut microbiome in treating patients with IBD.

Mechanism of Jianpi Qingchang Huashi Recipe in treating ulcerative colitis:A study based on network pharmacology and molecular docking

BACKGROUND Ulcerative colitis(UC)is a refractory intestinal disease with alternating onset and remission and a long disease course,which seriously affects the health and quality of life of patients.The goal of treatment is to control clinical symptoms,induce and maintain remission,promote mucosal healing,and reduce recurrence.Clinical trials have shown unsatisfactory clinical response rates.As a supplementary alternative medicine,traditional Chinese medicine has a rich history and has shown good results in the treatment of UC.Because of the quality of herbal medicine and other factors,the curative effect of traditional Chinese medicine is not stable enough.The mechanism underlying the effect of Jianpi Qingchang Huashi Recipe(JPQCHSR)on inducing UC mucosal healing is not clear.AIM To investigate the potential mechanism of JPQCHSR for the treatment of UC based on network pharmacology and molecular docking.METHODS Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform was used to extract the active components and action targets of JPQCHSR,and the target names were standardized and corrected through UniProt database.The related targets of UC were obtained through GeneCards database,and the intersection targets of drugs and diseases were screened by jvenn online analysis tool.The visual regulatory network of"Traditional Chinese medicine-active components-target-disease"was constructed using Cytoscape software,the protein interaction network was constructed using STRING database,and enrichment analysis of gene ontology and Kyoto Encyclopedia of Genes and Genomes pathways was conducted through R software.At last,the active components were docked with the core target through SYBYL-X 2.1.1 software.RESULTS Through database analysis,a total of 181 active components,302 targets and 205 therapeutic targets were obtained for JPQCHSR.The key compounds include quercetin,luteolin,kaempferol,etc.The core targets involved STAT3,AKT1,TP53,MAPK1,MAPK3,JUN,TNF,etc.A total of 2861 items were obtained by GO enrichment analysis,and 171 items were obtained by KEGG(Kyoto Encyclopedia of Genes and Genomes)pathway enrichment analysis.The results of molecular docking showed that the key active components in JPQCHSR had certain affinity with the core target.CONCLUSION The treatment of UC with JPQCHSR is a complex process of multi-component,multi-target and multi-pathway regulation.The mechanism of this Recipe in the treatment of UC can be predicted through network pharmacology and molecular docking,so as to provide theoretical reference for it to better play its therapeutic role.

Role of metabolites derived from gut microbiota in inflammatory bowel disease

Over the past two decades,it is improved gut microbiota plays an important role in the health and disease pathogenesis.Metabolites,small molecules produced as intermediate or end products of microbial metabolism,is considered as one of the major interaction way for gut microbiota with the host.Bacterial metabolisms of dietary substrates,modification of host molecules or bacteria are the major source of metabolites.Signals from microbial metabolites affect immune maturation and homeostasis,host energy metabolism as well as mucosal integrity maintenance.Based on many researches,the composition and function of the microbiota can be changed,which is also seen in the metabolite profiles of patients with inflammatory bowel disease(IBD).Additionally,some specific classes of metabolites also can trigger IBD.In this paper,definition of the key classes of microbialderived metabolites which are changed in IBD,description of the pathophysiological basis of association and identification of the precision therapeutic modulation in the future are the major contents.

Molecular regulation mechanism of intestinal stem cells in mucosal injury and repair in ulcerative colitis

Ulcerative colitis(UC)is a chronic nonspecific inflammatory disease with complex causes.The main pathological changes were intestinal mucosal injury.Leucinerich repeat-containing G protein coupled receptor 5(LGR5)-labeled small intestine stem cells(ISCs)were located at the bottom of the small intestine recess and inlaid among Paneth cells.LGR5+small ISCs are active proliferative adult stem cells,and their self-renewal,proliferation and differentiation disorders are closely related to the occurrence of intestinal inflammatory diseases.The Notch signaling pathway and Wnt/β-catenin signaling pathway are important regulators of LGR5-positive ISCs and together maintain the function of LGR5-positive ISCs.More importantly,the surviving stem cells after intestinal mucosal injury accelerate division,restore the number of stem cells,multiply and differentiate into mature intestinal epithelial cells,and repair the damaged intestinal mucosa.Therefore,in-depth study of multiple pathways and transplantation of LGR5-positive ISCs may become a new target for the treatment of UC.

Value of CT-guided core-needle biopsy in diagnosis and classification of malignant lymphomas using automated biopsy gun

AIM: To evaluate the value of CT-guided core-needle biopsy in diagnosis and classification of malignant lymphomas.METHODS: From January 1999 to October 2004, CT-guided core-needle biopsies were performed in 80 patients with suspected malignant lymphoma. Biopsies were performed with an 18-20 G biopsy-cut (CR Bard, Inc., Covington, GA,USA) needle driven by a spring-loaded Bard biopsy gun.RESULTS: A definite diagnosis and accurate histological subtype were obtained in 61 patients with a success rate of 76.25% (61/80). Surgical sampling was performed in 19 patients (23.75%) with non-diagnostic core-needle biopsies. The success rate of CT-guided core-needle biopsy varied with the histopathologic subtypes in our group.The relatively high success rates of core-needle biopsy were noted in diffuse large B-cell non-Hodgkin's lymphoma (NHL, 88.89%) and peripheral T-cell NHL (90%). However,the success rates were relatively low in anaplastic large cell (T/null cell) lymphoma (ALCL, 44.44%) and Hodgkin's disease (HD, 28.57%) in our group.CONCLUSION: CT-guided core-needle biopsy is a reliable means of diagnosing and classifying malignant lymphomas,and can be widely applied in the management of patients with suspected malignant lymphoma.

New insights into the interplay between intestinal flora and bile acids in inflammatory bowel disease

Intestinal flora plays a key role in nutrient absorption,metabolism and immune defense,and is considered to be the cornerstone of maintaining the health of human hosts.Bile acids synthesized in the liver can not only promote the absorption of fat-soluble substances in the intestine,but also directly or indirectly affect the structure and function of intestinal flora.Under the action of intestinal flora,bile acids can be converted into secondary bile acids,which can be reabsorbed back to the liver through the enterohepatic circulation.The complex dialogue mechanism between intestinal flora and bile acids is involved in the development of intestinal inflammation such as inflammatory bowel disease(IBD).In this review,the effects of intestinal flora,bile acids and their interactions on IBD and the progress of treatment were reviewed.

Jianpi Qingchang Bushen decoction improves inflammatory response and metabolic bone disorder in inflammatory bowel disease-induced bone loss

BACKGROUND Bone loss and osteoporosis are commonly described as extra-intestinal manifestations of inflammatory bowel disease(IBD).Jianpi Qingchang Bushen decoction(JQBD)is a prescription used in clinical practice.However,further studies are needed to determine whether JQBD regulates the receptor activator of nuclear factor kappa B(NF-κB)(RANK)/receptor activator of NF-κB ligand(RANKL)/osteoprotegerin(OPG)pathways and could play a role in treating IBD-induced bone loss.AIM To evaluate the therapeutic effect of JQBD in IBD-induced bone loss and explore the underlying mechanisms.METHODS An IBD-induced bone loss model was constructed by feeding 126-to-8-wk-old interleukin-10(IL-10)-knockout mice with piroxicam for 10 d.The mice were randomly divided into model and JQBD groups.We used wild-type mice as a control.The JQBD group was administered the JQBD suspension for 2 wk by gavage,while the control and model groups were given normal saline at the corresponding time points.All mice were killed after the intervention.The effect of JQBD on body weight,disease activity index(DAI),and colon length was analyzed.Histopathological examination,colon ultrastructure observation,and micro-computed tomographic scanning of the lumbar vertebrae were performed.The gene expression of NF-κB,tumor necrosis factor-α(TNF-α),IL-1β,IL-6,and IL-8 in the colon was evaluated by real-time polymerase chain reaction.Colon samples were assessed by Western blot for the expression of RANKL,OPG,RANK,and NF-κB proteins.RESULTS The model group lost body weight,had a shorter colon,and showed a dramatic increase in DAI score,whereas JQBD had protective and therapeutic effects.Treatment with JQBD significantly improved inflammatory cell infiltration and reduced crypt abscess and ulcer formation.Threedimensional imaging of the vertebral centrum in the model group revealed a lower bone mass,loose trabeculae,and“rod-shaped”changes in the structure compared to the control group and JQBD groups.The bone volume/total volume ratio and bone mineral density were significantly lower in the model group than in the control group.JQBD intervention downregulated the NF-κB,TNF-α,IL-1β,IL-6,and IL-8 m RNA expression levels.The RANKL and OPG protein levels were also improved.CONCLUSION JQBD reduces inflammation of the colonic mucosa and inhibits activation of the RANK/RANKL/OPG signaling pathway,thereby reducing osteoclast activation and bone resorption and improving bone metabolism.

Network pharmacology and molecular docking reveal zedoary turmeric-trisomes in Inflammatory bowel disease with intestinal fibrosis

BACKGROUND Inflammatory bowel disease(IBD) is a complex chronic IBD that is closely associated with risk factors such as environment, diet, medications and lifestyle that may influence the host microbiome or immune response to antigens. At present, with the increasing incidence of IBD worldwide, it is of great significance to further study the pathogenesis of IBD and seek new therapeutic targets. Traditional Chinese medicine(TCM) treatment of diseases is characterized by multiple approaches and multiple targets and has a long history of clinical application in China. The mechanism underlying the effect of zedoary turmerictrisomes on inducing mucosal healing in IBD is not clear.AIM To explore the effective components and potential mechanism of zedoary turmeric-trisomes in the treatment of IBD with intestinal fibrosis using network pharmacology and molecular docking techniques.METHODS The chemical constituents and targets of Rhizoma zedoary and Rhizoma sanarum were screened using the TCMSP database. The GeneCards database was searched to identify targets associated with intestinal fibrosis in IBD. The intersection of chemical component targets and disease targets was obtained using the Venny 2.1 online analysis platform, and the common targets were imported into the STRING 11.0 database to construct a protein interaction regulatory network. A “zedoary turmeric-trisomes-chemical composition-target-disease” network diagram was subsequently constructed using Cytoscape 3.7.2 software, and the topological properties of the network were analyzed using the “Network Analysis” plug-in. Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses of the common targets were performed using the DAVID 6.8 database to elucidate the mechanism of zedoary turmeric-trisomes in the treatment of IBD. Subsequently, molecular docking of the compounds and targets with the highest intermediate values in the “zedoary turmeric-trisomes-chemical composition-target-disease” network was performed using Sybyl-x 2.1.1 software.RESULTS A total of 5 chemical components with 60 targets were identified, as well as 3153 targets related to IBD and 44 common targets. The protein-protein interaction network showed that the core therapeutic targets included JUN, MAPK14, CASP3, AR, and PTGS2. The GO enrichment analysis identified 759 items, and the KEGG enrichment analysis yielded 52 items, including the cancer pathway, neuroactive ligand-receptor interaction, hepatitis B, and the calcium signaling pathway, reflecting the complex biological processes of the multicomponent, multitarget and multipathway treatment of diseases with zedoary turmeric-trisomes. Molecular docking showed that the compound bonded with the target through hydrogen bond interactions and exhibited good docking activity.CONCLUSION This study identified the potential mechanism of action of zedoary turmeric-trisomes in the treatment of inflammatory bowel fibrosis using network pharmacology and molecular docking technology, providing a scientific basis for further expansion of their clinical use.

Role of adherent invasive Escherichia coli in pathogenesis of inflammatory bowel disease

Gut microbiota imbalances play an important role in inflammatory bowel disease(IBD),but no single pathogenic microorganism critical to IBD that is specific to the IBD terminal ileum mucosa or can invade intestinal epithelial cells has been found.Invasive Escherichia coli(E.coli)adhesion to macrophages is considered to be closely related to the pathogenesis of inflammatory bowel disease.Further study of the specific biological characteristics of adherent invasive E.coli(AIEC)may contribute to a further understanding of IBD pathogenesis.This review explores the relationship between AIEC and the intestinal immune system,discusses the prevalence and relevance of AIEC in Crohn's disease and ulcerative colitis patients,and describes the relationship between AIEC and the disease site,activity,and postoperative recurrence.Finally,we highlight potential therapeutic strategies to attenuate AIEC colonization in the intestinal mucosa,including the use of phage therapy,antibiotics,and anti-adhesion molecules.These strategies may open up new avenues for the prevention and treatment of IBD in the future.

Clinicopathological features and expression of regulatory mechanism of the Wnt signaling pathway in colorectal sessile serrated adenomas/polyps with different syndrome types

BACKGROUND Colorectal cancer(CRC)is the third most common cancer worldwide,with the fourth highest mortality among all cancers.Reportedly,in addition to adenomas,serrated polyps,which account for 15%-30%of CRCs,can also develop into CRCs through the serrated pathway.Sessile serrated adenomas/polyps(SSAs/Ps),a type of serrated polyps,are easily misdiagnosed during endoscopy.AIM To observe the difference in the Wnt signaling pathway expression in SSAs/Ps patients with different syndrome types.METHODS From January 2021 to December 2021,patients with SSAs/Ps were recruited from the Endoscopy Room of Shanghai Traditional Chinese Medicine-Integrated Hospital,affiliated with Shanghai University of Traditional Chinese Medicine.Thirty cases each of large intestine damp-heat(Da-Chang-Shi-Re,DCSR)syndrome and spleen-stomach weakness(Pi-Wei-Xu-Ruo)syndrome were reported.Baseline comparison of the general data,typical tongue coating,colonoscopy findings,and hematoxylin and eosin findings was performed in each group.The expression of the Wnt pathway-related proteins,namelyβ-catenin,adenomatous polyposis coli,and mutated in colorectal cancer,were analyzed using immunohistochemistry.RESULTS Significant differences were observed with respect to the SSAs/Ps size between the two groups of patients with different syndrome types(P=0.001).The other aspects did not differ between the two groups.The Wnt signaling pathway was activated in patients with SSAs/Ps belonging to both groups,which was manifested asβ-catenin protein translocation into the nucleus.However,SSAs/Ps patients with DCSR syndrome had more nucleation,higherβ-catenin expression,and negative regulatory factor(adenomatous polyposis coli and mutated in colorectal cancer)expression(P<0.0001)than SSA/P patients with Pi-Wei-Xu-Ruo syndrome.In addition,the SSA/P size was linearly correlated with the related protein expression.CONCLUSION Patients with DCSR syndrome had a more obvious Wnt signaling pathway activation and a higher risk of carcinogenesis.A high-quality colonoscopic diagnosis was essential.The thorough assessment of clinical diseases can be improved by combining the diseases of Western medicine with the syndromes of traditional Chinese medicine.

Research progress on the relationship between Paneth cellssusceptibility genes,intestinal microecology and inflammatory bowel disease

Inflammatory bowel disease(IBD)is a disorder of the immune system and intestinal microecosystem caused by environmental factors in genetically susceptible people.Paneth cells(PCs)play a central role in IBD pathogenesis,especially in Crohn's disease development,and their morphology,number and function are regulated by susceptibility genes.In the intestine,PCs participate in the formation of the stem cell microenvironment by secreting antibacterial particles and play a role in helping maintain the intestinal microecology and intestinal mucosal homeostasis.Moreover,PC proliferation and maturation depend on symbiotic flora in the intestine.This paper describes the interactions among susceptibility genes,PCs and intestinal microecology and their effects on IBD occurrence and development.

单发肝细胞癌合并微血管侵犯患者根治性切除术后辅助经导管动脉化疗栓塞治疗的疗效和安全性:一项随机临床试验

背景与目的单发肝细胞癌(hepatocellular carcinoma,HCC)合并微血管侵犯(microvascular invasion,MVI)患者根治性切除术后的最佳辅助治疗方案一直存有争议。本试验旨在评估肝切除术后辅助经导管动脉化疗栓塞(transcatheter arterial chemoembolization,TACE)与单纯肝切除术对直径≥5 cm单发HCC合并MVI患者的疗效和安全性。方法在本随机、开放性、III期试验中,将直径≥5 cm单发HCC合并MVI患者随机分为2组(1∶1):在肝切除术后接受1–2个周期的辅助TACE治疗(肝切除–TACE组)或单纯接受肝切除(单纯肝切除组)。主要终点是无病生存期(disease-free survival,DFS),次要终点包括总生存期(overall survival,OS)和不良事件。结果在2009年6月1日至2012年12月31日期间,共纳入250例患者,随机分为肝切除–TACE组(n=125)或单纯肝切除组(n=125)。两组患者的临床病理特征相似。从随机开始的中位随访时间为37.5个月(四分位距为18.3–48.2个月)。肝切除–TACE组的中位DFS显著长于单纯肝切除组[17.45个月(95%置信区间,confidence interval,CI:11.99–29.14)vs. 9.27个月(95%CI:6.05–13.70),风险比(hazard ratio,HR)=0.70(95%CI:0.52–0.95),P=0.020]。肝切除–TACE组中位OS也显著长于单纯肝切除组[44.29个月(95%CI:25.99–62.58)vs. 22.37个月(95%CI:10.84–33.91),HR=0.68(95%CI:0.48–0.97),P=0.029]。治疗相关不良事件在肝切除–TACE组中更为多见,虽然这些不良事件一般都是轻度和可控的。两组中最常见的3级或4级不良事件为中性粒细胞减少和肝功能异常。结论对于直径≥5 cm单发HCC合并MVI患者,根治术后进行辅助TACE治疗是一种合适的选择,且毒性是可接受的。

Single-cell RNA-sequencing combined with bulk RNA-sequencing analysis of peripheral blood reveals the characteristics and key immune cell genes of ulcerative colitis

BACKGROUND Ulcerative colitis(UC)is a complicated disease caused by the interaction between genetic and environmental factors that affects mucosal homeostasis and triggers an inappropriate immune response.Single-cell RNA sequencing(scRNA-seq)can be used to rapidly obtain the precise gene expression patterns of thousands of cells in the intestine,analyze the characteristics of cells with the same phenotype,and provide new insights into the growth and development of intestinal organs,the clonal evolution of cells,and immune cell changes.These findings can provide new ideas for the diagnosis and treatment of intestinal diseases.To identify clinical phenotypes and biomarkers that can predict the response of UC patients to specific therapeutic drugs and thus aid the diagnosis and treatment of UC.METHODS Using the Gene Expression Omnibus(GEO)database,we analyzed peripheral blood cell subtypes of patients with UC by scRNA-seq combined with bulk RNA sequencing(RNA-seq)to reveal the core genes of UC.We then combined weighted gene correlation network analysis(WGCNA)and least absolute shrinkage and selection operator(LASSO)analysis to reveal diagnostic markers of UC.RESULTS After processing the scRNA-seq data,we obtained data from approximately 24340 cells and identified 17 cell types.Through intercellular communication analysis,we selected monocyte marker genes as the candidate gene set for the prediction model.Construction of a WGCNA coexpression network identified RhoB,cathepsin D(CTSD)and zyxin(ZYX)as core genes.Immune infiltration analysis showed that these three core genes were strongly correlated with immune cells.Functional enrichment analysis showed that the differentially expressed genes were closely related to immune and inflammatory responses,which are associated with many challenges in the diagnosis and treatment of UC.CONCLUSION Through scRNA-seq analysis,LASSO diagnostic model building and WGCNA,we identified RhoB,CTSD and ZYX as core genes of UC that are closely related to monocyte infiltration that may serve as diagnostic markers and molecular targets for UC therapeutic intervention.

Jianpi Qingchang Decoction-containing serum regulates the autophagy ofinterstitial cells

Objective： To investigate the effects of Jianpi Qingchang Decoction-containing serum （JQD-CS） on interstitial cells ofCajal （ICCs） autophagy and cell cycle arrest in vitro. Methods： ICCs were collected from the small intestines of miceand analyzed using an anti-c-Kit antibody. ICCs were divided into five groups： the blank group, the rapamycin group （anautophagy inducer）, the 5% （rapamycin ＋ 5% JQD-CS）, the 20% （rapamycin ＋ 20% JQD-CS） JQD-CS groups, and the3-Methyladenine （3-MA） group （rapamycin ＋ 3-MA; positive control）. Transmission electron microscopy was used toobserve the ultrastructure of ICCs. Western blotting was used to detect the expression of microtubule-associated protein1 light chain 3 （LC3-II）, Beclin-1, phosphatidylinositol 3-kinase （PI3K）, p-PI3K, protein kinase B （AKt）, p-AKt,mammalian target of rapamycin （mTOR）, and p-mTOR. Ca2＋ current was examined by patch-clamp experiments. Cellcycle was detected by flow cytometry. Results： Unlike in the rapamycin group, the ICC structures were more integratedand a lower number of autophagic vacuoles were observed in the 20% JQD-CS group. Moreover, the expression ofLC3-II and Beclin-1 decreased, the expression of c-Kit, p-PI3K, p-AKt increased, the maximum current density valuedecreased, and the number of cells in the G1 phase increased while those in the G2/M phase decreased, with the additionof 20% JQD-CS. Conclusion： JQD-CS can antagonize rapamycin-induced autophagy in ICCs in vitro by promoting thephosphorylation of PI3K/AKt pathway, inhibiting Ca2 ＋ inflow, and regulating the cell cycle.

基于“脾虚-内质网应激”探讨健脾法对炎症性肠病肠纤维化内质网应激的调节作用

肠纤维化是炎症性肠病(inflammatory bowel disease,IBD)常见并发症之一,内质网应激参与纤维化形成是其重要发病机制.内质网是细胞内最大的膜网络结构,主要参与机体蛋白质的合成代谢.脾主运化是脾脏正常的生理功能,脾虚不能运化水谷精微,导致湿热、痰浊、瘀毒等病理产物壅滞于肠道,肠壁增厚,甚至肠腔狭窄、梗阻,诱发本病.生理学上,脾和内质网有一定相关性;病理学上,脾虚是内质网应激的本质,是IBD肠纤维化的核心病理环节.因此,从健脾法治疗有助于改善内质网代谢功能.所以,基于“脾虚-内质网应激”理论探讨健脾法调节内质网应激、恢复内质网稳态是研究和治疗IBD肠纤维化作用机制的重要理论.

Adjuvant transcatheter arterial chemoembolization after curative resection for hepatocellular carcinoma patients with solitary tumor and microvascular invasion: a randomized clinical trial of efficacy and safety

Background:The optimal strategy for adjuvant therapy after curative resection for hepatocellular carcinoma(HCC)patients with solitary tumor and microvascular invasion(MVI)is controversial.This trial evaluated the efficacy and safety of adjuvant transcatheter arterial chemoembolization(TACE)after hepatectomy versus hepatectomy alone in HCC patients with a solitary tumor≥5 cm and MVI.Methods:In this randomized,open-labeled,phase III trial,HCC patients with a solitary tumor≥5 cm and MVI were randomly assigned(1:1)to receive either 1-2 cycles of adjuvant TACE after hepatectomy(Hepatectomy-TACE)or hepatectomy alone(Hepatectomy Alone).The primary endpoint was disease-free survival(DFS);the secondary end-points included overall survival(OS)and adverse events.Results:Between June 1,2009,and December 31,2012,250 patients were enrolled and randomly assigned to the Hepatectomy-TACE group(n=125)or the Hepatectomy Alone group(n=125).Clinicopathological characteristics were balanced between the two groups.The median follow-up time from randomization was 37.5 months[interquartile range 18.3-48.2 months].The median DFS was significantly longer in the Hepatectomy-TACE group than in the Hepatectomy Alone group[17.45 months(95%confidence interval[CI]11.99-29.14)vs.9.27 months(95%CI 6.05-13.70),hazard ratio[HR]=0.70(95%CI 0.52-0.95),P=0.020],respectively.The median OS was also significantly longer in the Hepatectomy-TACE group than in the Hepatectomy Alone group[44.29 months(95%CI 25.99-62.58)vs.22.37 months(95%CI 10.84-33.91),HR=0.68(95%CI 0.48-0.97),P=0.029].Treatment-related adverse events were more frequently observed in the Hepatectomy-TACE group,although these were generally mild and manageable.The most common grade 3 or 4 adverse events in both groups were neutropenia and liver dysfunction.Conclusion:Hepatectomy followed by adjuvant TACE is an appropriate option after radical resection in HCC patients with solitary tumor≥5 cm and MVI,with acceptable toxicity.