ObjectiveTo study on the different therapeutic effects and potential mechanisms of Rhei Radix et Rhizoma（RH） before and after steaming with wine on constipation model mice. MethodsFifty-four male ICR mice were randomly divided into control group， model group， lactulose group（1.5 mg·kg^-1）， high， medium and low dose groups of RH and RH steaming with wine（PRH）（8， 4， 1 g·kg^-1）. Except for the control group， the constipation model was replicated by gavage of loperamide hydrochloride（6 mg·kg^-1） in the other groups. After 2 weeks of modeling， each administration group was gavaged with the corresponding dose of drug solution， and the control and model groups were given an equal volume of normal saline， 1 time/d for 2 consecutive weeks. After administration， the feces were collected for 16S rRNA sequencing， the levels of gastrin（GAS）， motilin（MTL）， interleukin-6（IL-6）， γ-interferon（IFN-γ） in the colonic tissue were detected by enzyme-linked immunosorbent assay（ELISA）， the histopathological changes of colon were observed by hematoxylin-eosin（HE） staining， flow cytometry was used to detect the proportion changes of CD4⁺， CD8⁺ and regulatory T cell（Treg） in peripheral blood. ResultsCompared with the control group， the model group showed significantly decrease in fecal number in 24 h， fecal quality and fecal water rate（P<0.01）， the colon was seen to have necrotic shedding of mucosal epithelium， localized intestinal glands in the lamina propria were degenerated， necrotic and atrophied， a few lymphocytes were seen to infiltrate in the necrotic area in a scattered manner， the contents of GAS and MTL， the proportions of CD4⁺， CD8⁺ and Treg were significantly reduced（P<0.01）， the contents of IL-6 and IFN-γ were significantly elevated（P<0.05， P<0.01）. Compared with the model group， the fecal number in 24 h， fecal quality and fecal water rate of high-dose groups of RH and PRH were significantly increased（P<0.05， P<0.01）， the pathological damage of the colon was alleviated to varying degrees， the contents of GAS， MTL， IL-6 and IFN-γ were significantly regressed（P<0.05， P<0.01）， and the proportions of CD4⁺ and CD8⁺ were significantly increased（P<0.01）， although the proportion of Treg showed an upward trend， there was no significant difference. In addition， the results of intestinal flora showed that the number of amplicon sequence variant（ASV） and Alpha diversity were decreased in the model group compared with the control group， and there was a significant difference in Beta diversity， with a decrease in the relative abundance of Lactobacillus and an increase in the relative abundances of Bacillus and Helicobacter. Compared with the model group， the ASV number and Alpha diversity were increased in the high-dose groups of RH and PRH， and there was a trend of regression of Beta diversity to the control group， the relative abundance of Lactobacillus increased， and the relative abundances of Bacillus and Helicobacter decreased. ConclusionRH and PRH can improve dysbacteriosis， promote immune system activation， inhibit the release of inflammatory factors for enhancing the gastrointestinal function， which may be one of the potential mechanisms of their therapeutic effect on constipation.

Dihydroorotate dehydrogenase (DHODH) is a flavin-dependent metabolic enzyme that oxidizes dihydroorotate acid to orotic acid in the de novo synthesis pathway of pyrimidine metabolism. DHODH is located in mitochondria, closely related to cellular oxidative phosphorylation, and an important suppressor of the ferroptosis pathway. This study investigates the influence of DHODH on the progression of malignant tumors, including its important role in the de novo synthesis of pyrimidine, oxidative phosphorylation, and ferroptosis. The objective is to present evidence that DHODH is a potential target for the clinical treatment of tumors.

Objective:To investigate the correlation factors of complete clinical response in idiopathic inflammatory myopathies(IIMs)patients receiving conventional treatment.Methods:Patients diagnosed with IIMs hospitalized in Peking University People's Hospital from January 2000 to June 2023 were in-cluded.The correlation factors of complete clinical response to conventional treatment were identified by analyzing the clinical characteristics,laboratory features,peripheral blood lymphocytes,immunological indicators,and therapeutic drugs.Results:Among the 635 patients included,518 patients finished the follow-up,with an average time of 36.8 months.The total complete clinical response rate of IIMs was 50.0％(259/518).The complete clinical response rate of dermatomyositis(DM),anti-synthetase syn-drome(ASS)and immune-mediated necrotizing myopathy(IMNM)were 53.5％,48.9％and 39.0％,respectively.Fever(P=0.002)and rapid progressive interstitial lung disease(RP-ILD)(P=0.014)were observed much more frequently in non-complete clinical response group than in complete clinical re-sponse group.The aspartate transaminase(AST),lactate dehydrogenase(LDH),D-dimer,erythrocyte sedimentation rate(ESR),C-reaction protein(CRP)and serum ferritin were significantly higher in non-complete clinical response group as compared with complete clinical response group.As for the treat-ment,the percentage of glucocorticoid received and intravenous immunoglobin(IVIG)were significantly higher in non-complete clinical response group than in complete clinical response group.Risk factor analysis showed that IMNM subtype(P=0.007),interstitial lung disease(ILD)(P=0.001),eleva-ted AST(P=0.012),elevated serum ferritin(P=0.016)and decreased count of CD4+T cells in peripheral blood(P=0.004)might be the risk factors for IIMs non-complete clinical response.Conclu-sion:The total complete clinical response rate of IIMs is low,especially for IMNM subtype.More effec-tive intervention should be administered to patients with ILD,elevated AST,elevated serum ferritin or decreased count of CD4+T cells at disease onset.

Objective:To analyze the efficacy of anterior cervical Hybrid surgery and posterior cervical expansive open-door laminoplasty(EODL)in the treatment of multilevel cervical spondylotic myelopathy,and to discuss the selection of surgical methods for the patients with multilevel cervical spondylotic myelopathy.Methods:The retrospective analysis was conducted of 70 patients with multilevel cervical spondylotic myelopathy who underwent surgery at Affilated Beijing Traditional Chinese Medicine Hospital of Capital Medical University from July 2017 to July 2020.Based on the different surgical methods,the patients were divided into anterior group(n=35)and posterior group(n=35).The patients in anterior group underwent Hybrid surgery[anterior cervical discectomy and fusion(ACDF)combined with artificial cervical disc replacement(ACDR)],and the patients in posterior group underwent EODL.The hospitalization time,operation time,intraoperative blood loss,and postoperative drainage volume of the patients in two groups were recorded;the efficacy was evaluated by Japanese orthopaedic association(JOA)score,JOA improvement rate,neck disability index(NDI),visual analogue scale(VAS)for pain,and postoperative satisfaction score;the complications of the patients in two groups after surgery were recorded.Results:Compared with posterior group,the intraoperative blood loss,postoperative drainage volume,hospitalization time,and operation time of the patients in anterior group were significantly decreased(P<0.01),and the preoperative score had no significant difference(P>0.05).At the final follow-up after surgery,compared with posterior group,the JOA score and JOA improvement rate of the patients in anterior group were significantly increased(P<0.01),and the NDI score and VAS score were significantly decreased(P<0.01).Compared with before surgery,the JOA scores of the patients in two groups at the final follow-up after surgery were increased(P<0.01),and the NDI and VAS scores were significant decreased(P<0.01).The postoperative satisfaction of the patients in two groups was high based on the postoperative satisfaction score.There was no significant difference in the incidence of postoperative complication of the patients between two groups(P>0.05).Conclusion:Both the anterior cervical Hybrid surgery and EODL achieve the satisfactory results in the treatment of multilevel cervical spondylotic myelopathy.Hybrid surgery has the advantages of less bleeding and shorter surgery time,and the most suitable surgical method should be chosen clinically based on the actual situation of the patients.

Improving the reproducibility of biomedical research results is a major challenge. Transparent and accurate reporting of the research process enables readers to evaluate the reliability of the research results and further explore the experiment by repeating it or building upon its findings. The ARRIVE 2.0 guidelines, released in 2019 by the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs), provide a checklist that is applicable to any in vivo animal research report. These guidelines aim to improve the standardization of experimental design, implementation, and reporting, as well as enhance the reliability, repeatability, and clinical translation of animal experimental results. The use of the ARRIVE 2.0 guidelines not only enriches the details of animal experimental research reports, ensuring that information on animal experimental results is fully evaluated and utilized, but also enables readers to understand the content expressed by the author accurately and clearly, promoting the transparency and completeness of the fundamental research review process. At present, the ARRIVE 2.0 guidelines have been widely adopted by international biomedical journals. This article is based on the best practices following the ARRIVE 2.0 guidelines in international journals, and it interprets, explains, and elaborates in Chinese the fifth part of the comprehensive version of the ARRIVE 2.0 guidelines published in PLoS Biology in 2020 (the original text can be found at https://arriveguidelines.org). This section includes the items 6-11 of Recommended 11 section, covering "Animal Care and Monitoring", "Interpretation/Scientific Implications", "Generalisability/Translation", "Protocol Registration", "Data Access" and "Declaration of Interests". Its aim is to promote a comprehensive understanding and use of the ARRIVE 2.0 guidelines among domestic researchers, to enhance the standardization of experimental animal research and reporting, and to promote high-quality development of experimental animal sciences and comparative medicine research in China.

Objective:To screen the host interaction proteins of the severe fever with thrombocytopenia syndrome virus(SFTSV)nonstructural protein(NSs)by immunoprecipitation combined with mass spectrometry analysis,to discuss the functions,subcellular localization,and biological pathways of these interaction proteins,and to provide the basis for clarifying the replication and pathogenic mechanism of SFTSV.Methods:The eukaryotic expression vectors pSFTSV-NSs-Flag(experimental group)and Flag-CMV-3(negative group)were transfected into the human embryonic kidney 293T cells,and contorl group(no treatment)was set up.The lysates of the cells in various groups were collected,and the expression and localization of SFTSV NSs in the host cells were verified by indirect immunofluorescence and Western blotting methods.The protein lysates were treated with protein A/G and immunoprecipitation was used to enrich host proteins binding to NSs.The captured interaction proteins were initially analyzed by silver staining and Coomassie brilliant blue staining to observe the differential protein bands in various groups;liquid chromatography-tandem mass spectrometry was used to obtain the information of protein sequences;the reliable proteins were retained and searched by UniProt database;Gene Ontology(GO)functional enrichment analysis,IPR,eukaryotic orthologous groups(KOGs)functional annotation,Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis,subcellular localization,and transcription factor(TF)functional annotation were used to determine the subcellular structure,gene functions,and biological processes of the interaction proteins.Results:The immunofluorescence results showed that the SFTSV NSs expressed a single specific band at relative molecular mass 33 000 and was localized in the cytoplasm in a granular inclusion body-like manner.The silver staining and Coomassie brilliant blue staining results showed there were significant differential protein bands between experimental group and negative group.The mass spectrometry results identified 46 potential interaction proteins.The GO functional enrichment analysis,KOGs functional annotation,and KEGG signaling pathway enrichment analysis results showed that the biological pathways related to viral translation,cellular metabolism,and protein transport were enriched with a considerable number of proteins.Eight annotated proteins had intermediate filament domains.The highest percentage of subcellular localization was cytoplasmic proteins,consistent with the NSs localization site.The TF functional annotation analysis results showed one protein from the NF-Y family.Conclusion:The interaction proteins play roles in assisting the proper protein folding,participating in the cribosome translation,and forming the cytoskeleton,which may be involved in antiviral replication.These proteins can be used as candidate proteins for further study on the replication mechanism of SFTSV.

Objective:To discuss the host proteins that interact with the Nelson Bay orthoreovirus(NBV)σNS protein in the fibroblasts L929 of the mice,and to clarify the effect of host proteins on the viral replication.Methods:The bait plasmid pGBKT7-S3 expressing σNS protein was constructed,and sequencing technology was used to verify the accurate expression of the bait plasmid in the Y2H yeast cells.The pGBKT7-S3 and pGADT7 plasmids were separately and jointly transformed into the Y2HGold yeast cells,plated on solid medium,and the colony growth was observed to confirm that the σNS protein was non-toxic to the yeast cells and could not self-activate the reporter gene.The bait plasmid pGBKT7-S3 was hybridized with the cDNA library in fibroblasts L929 of the mice,and the plasmids encoding the interacting proteins were extracted from the positive clones.The positive sequencing results were screened for the proteins interacting with NBV σNS through the Uniprot database.Gene Ontology(GO)functional enrichment analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis,and STRING bioinformatics analysis were performed on the interacting proteins.Results:A total of 61 positive clones were successfully screened.The DNA sequencing analysis and BLAST alignment removed 23 positive clones that did not match the database or were similar in sequence.The positive sequencing results identified 38 proteins interacting with NBV σNS through the Uniprot database.Thirty-one proteins were involved in cellular biological processes;thirty-six proteins were cellular anatomical components;thirty-one proteins had binding functions.Five proteins were part of the mitochondrial respiratory chain;seven proteins were ribosomal proteins and components of the ribosomal subunits;two proteins were involved in iron metabolism homeostasis.The GO functional enrichment analysis results showed that the interacting proteins were enriched in biological processes(BP)such as cellular processes,metabolic processes,biological regulation,localization,and response to stimuli;the cellular components were mainly cellular anatomical components and protein-containing complexes;the molecular functions were concentrated in binding,catalytic activity,structural molecule activity,and transporter activity.The KEGG signaling pathway enrichment analysis results showed that the proteins were highly enriched in translation,folding,sorting,and degradation pathways of genetic information processing and were mainly associated with the digestive system in the organism;they were linked to various viral infections and cancers.The STRING analysis results showed that the interacting proteins included ribosomal proteins,protein modification proteins,metabolic proteins,and immune proteins.Conclusion:The host proteins that interact with NBV σNS protein are successfully screened,and these host proteins play important roles in viral replication.

Objective:To investigate the feasibility of conducting active surveillance (AS) for low risk papillary thyroid microcarcinoma (PTMC) in China and to examine the factors in association with disease progression during AS.Methods:This study was a prospective observational research conducted from Jun. 2018 to Aug. 2022 at Ruijin Hospital affiliated to Shanghai Jiao Tong University School of Medicine. Seventy-three patients with cytologically confirmed low-risk PTMC were enrolled in this study. They were followed up by ultrasonography, and the observed nodules were re-assessed whether or not to have disease progression. Disease progression was defined as having nodule enlarged more than 3 mm in any of diameters measured on ultrasound, or/and presence of suspicious lymph node metastasis.Results:The median follow-up time was 33 months. At the time of last follow-up, 16 cases (21.9%) exhibited disease progression, including 9 cases (12.3%) with suspicious lymph nodes detected by ultrasound, and 8 cases (11.0%) with lesion enlargement; one case (1.3%) exhibited both situations. The univariate chi-square analysis revealed that young patients (≤45 years old, P=0.041), presence of microcalcifications ( P=0.032), initial larger nodule (diameter greater than 7 mm, P=0.003), and elevated thyroid autoantibody levels ( P=0.008) were associated with disease progression. Multiple regression analysis showed elevated thyroid autoantibodies ( OR=4.311, P=0.030) and initial larger nodule ( OR=6.196, P=0.034) were independent risk factors for PTMC progression,respectively. Conclusions:AS for low-risk PTMC is a feasible and effective. During the observation, ultrasound can reveal suspicious lymph nodes and nodule enlargement, which are crucial indicators for assessing disease progression. Patients with initially larger nodule size and elevated thyroid autoantibody level are more likely to exhibit disease progression and should receive closer attention.

Kirsten rat sarcoma viral oncogene homolog(KRAS)is a type of gene closely related to human tumors.And it's an important medical index to access the tumor development,prognosis and the efficacy of chemoradiotherapy.RAS mutations,in which KRAS mutations account for up to 85%,are the most common oncogenic driving mutations in human tumors.The most frequent KRAS mutation sites are codons 12,13,61 and 146.Codon G12,as the most frequently mutated one,can be divided into multiple subtypes,with G12D mutation being the most common,followed by G12V,G12C,etc.Colorectal cancer(CRC)is one of the tumors with the highest frequency of KRAS mutations.Both G12D and G12V are the most common mutation subtypes in CRC.In the field of treatments for CRC with KRAS mutations,targeted therapy had not been possible until the release of KRASG12C inhibitors in 2013,and new drugs have been developed one after another since then.This study summarized the mutations of KRAS and the advances in clinical research,including the latest advances in targeted drugs,chemotherapy drugs,immunotherapy drugs,ferroptosis,and other treatment methods.Among them,in terms of targeted drugs,this review explored KRASG12C inhibitors(sotorasib,adagrasib,D-1553,IBI351,etc.),anti-angiogenic drugs(monoclonal antibodies such as bevacizumab,remdesizumab,etc),small molecule multi-target tyrosine kinase inhibitors such as sunitinib,etc.In terms of immunotherapy drugs,there have also been many advances,such as the ARETHUSA clinical trial,which found that temozolomide reduced the tumor mutational burden(TMB)of O-6-methylguanine-DNA methyltransferase(MGMT)deficiency and RAS mutation in patients with advanced metastatic colorectal cancer(mCRC),providing innovative ideas for patient immunotherapy.For example,the combination of xindilimab with bevacizumab,oxaliplatin,and capecitabine can be used for first-line treatment of RAS mutations,microsatellite stability(MSS),and unresectable mCRC.Relevant studies have shown that the combination therapy has good therapeutic potential and controllable tolerability safety.This review explored the mechanisms of KRAS mutations and the latest advances in clinical research and treatment,in order to provide reference for the treatment of KRAS mutated colorectal cancer.

Objective:To explore the mechanism of EPO on activating receptors of NK cells in rats with diabetic nephropathy based on HMGB1/Beclin-1 signaling pathway.Methods:Forty SPF male SD rats were randomly divided into normal group(A),model group(B),metformin group(C)and EPO group(D),with 10 rats in each group.Diabetic nephropathy modeling was performed on groups B,C and D using high-fat diet and streptozotocin.After successful modeling,group C was given metformin 100 mg/kg by intra-gastric administration,and group D was intraperitoneal injection of EPO 100 U/kg.Group A and B were given normal saline at the same volume by intragastric administration simultaneously.After successful modeling,the remaining 20 rats were randomly divided into group E and group F.Group E was given 30 mg/kg HMGB1 inhibitor by intragastric administration,group F was given 30 mg/kg HMGB1 inhibitor by intragastric administration and 100 U/kg EPO by intraperitoneal injection.HK-2 cells were taken and divided into high glucose+HK-2 cells(HH)group,EPO+high glucose+HK-2 cells(EH)group,glycyrrhizin L+high glucose+HK-2 cells(LH)group,and cultured with glucose for cell experiment.Blood glucose was detected by glucose analyzer,biochemical indexes were de-tected by automatic biochemical analyzer,and renal pathological morphology of rats was detected by PAS staining.Expression of HMGB1/Beclin-1 protein was detected by Western blot,and expressions of NK cell activated receptors were detected by RT-PCR and immunohistochemistry.Results:Compared with group A,blood glucose,24 h urine volume,blood glucose curve,BUN,Scr,UAlb contents in blood and urine,mRNA and protein expressions of NKp30,NKp44,NKp46 in group B were significantly increased(P<0.05),while body weight was significantly decreased(P<0.05).Compared with group B,blood glucose,24 h urine volume,blood glucose curve,BUN,Scr,UAlb contents in blood and urine,mRNA and protein expression of NKp30,NKp44 and NKp46 in group C and group D were significantly decreased(P<0.05),while body weight was significantly increased(P<0.05),renal pathology was also significantly improved,the changes of group D was significantly higher than that of group C(P<0.05).Compared with group A,expression of HMGB1/Beclin-1 protein in renal tissues of rats in group B was significantly increased(P<0.05),and expression of HMGB1/Beclin-1 protein in renal tissues of rats in groups C,D,E and F was significantly decreased(P<0.05),expression of HMGB1/Beclin-1 protein in group D was significantly decreased compared with group C(P<0.05),there was no significant difference between group E and group D,and expression of HMGB1/Beclin-1 protein in group F was significantly decreased compared with group E.HMGB1/Beclin-1 protein expression was positively correlated with NKp30,NKp44 and NKp46 mRNA(P<0.05).Compared with HH group,proliferation rates of EH and LH groups were significantly increased(P<0.05),while apoptosis rate and HMGB1/Beclin-1 protein expression were significantly decreased(P<0.05),there was no significant difference between LH group and EH group(P>0.05).Conclusion:EPO can effectively reduce expressions of NK cell activated receptors and inhibit HMGB1/Beclin-1 signaling path-way in diabetic nephropathy rats,suggesting that EPO may exert its effect by regulating NK cell activated receptors and HMGB1/Beclin-1 signaling pathway.NK cell activation receptors expressions are positively correlated with HMGB1/Beclin-1 signaling pathway.