The rationale for the design of control groups in tuina clinical trial is the foundation for rigorously validating the effectiveness and safety of this therapy. This article reviewed the current state of the design of tuina placebo in control groups of clinical trials， pointed out the necessity of setting up tuina placebo in clinical trials of tuina， analyzed the challenges in implementing blinding of tuina manipulation， and concluded that tuina placebo is still challenged by the placebo effect， the diversification of tuina manipulation but the lack of standardization， and the difficulty of implementing blinding due to the high level of public awareness of tuina. This article also summarized the design of placebo manipulation in three types of clinical trials， including spinal manipulation， acupressure， and paediatric tuina， and proposed four strategies for designing placebo tuina manipulation-controlling placebo effects， developing operational standards for placebo tuina manipulation， ensuring the rigor of blinding implementation， and applying new technologies to enhance the standardization and blinding capacity of placebo tuina methods. So the article is aimed at improving the methodological quality of tuina clinical trial designs， and promoting the standardization and scientificity of tuina clinical trial design.

Objective To study the current status of the randomized controlled trial(RCT)outcome indicators of acupuncture and moxibustion in the treatment of chronic fatigue syndrome(CFS);To provide guidance and references for clinical research and protocol design.Methods RCT of acupuncture and moxibustion for CFS were retrieved from CNKI,Wanfang Data,VIP,SinoMed,PubMed,Embase,Web of Science databases from January 1,2015 to December 9,2024.Through literature screening and data extraction,Excel 2019 was used to make a descriptive analysis of the basic characteristics,diagnostic criteria,intervention measures,classification of outcome indicators,use frequency and evaluation time points of the included studies.Origin2021 software was used to draw bubble charts for visual display.Results Totally 119 RCT were included,involving 8 445 subjects,5 diagnostic criteria of Western medicine,5 diagnostic criteria of TCM,and 6 outcome indicators,namely clinical symptoms/signs,symptoms/syndrome of TCM,physical and chemical examination,quality of life,safety evaluation,and patient satisfaction.A total of 58 outcome indicators were reported,with a cumulative frequency of 430 times.Conclusion The RCT of acupuncture and moxibustion treatment of CFS have some problems,such as incomplete baseline assessment,lack of sample size calculation basis,high risk of bias,ignorance of primary and secondary indicators in the selection of outcome indicators,numerous indicators selection and measurement points,lack of long-term follow-up,inconsistent evaluation criteria for TCM syndrome efficacy,and insufficient attention to health economics evaluation and safety outcome indicators.

As a biocatalyst, laccase has been widely studied and applied in the papermaking industry. However, the low catalytic efficiency and poor stability of natural laccase limit its application in the pulping process. To develop the laccase with high activity and strong tolerance, we carried out directed evolution for modification of the laccase derived from Bacillus pumilus and screened out the mutants F282L/F306L and Q275P from the random mutant library by high-throughput screening. The specific activities of F282L/F306L and Q275P were 280.87 U/mg and 453.94 U/mg, respectively, which were 1.42 times and 2.30 times that of the wild-type laccase. Q275P demonstrated significantly improved thermal stability, with the relative activity 20% higher than that of the wild-type laccase after incubation at 40 ℃, 50 ℃, and 70 ℃ for 4 h. F282L/F306L and Q275P showed greater tolerance to metal ions and organic solvents than the wild-type laccase. The K_m value of the wild-type laccase was 374.97 μmo/L, and those of F282L/F306L and Q275P were reduced to 318.96 μmo/L and 360.71 μmo/L, respectively, which suggested that the substrate affinity of laccase was improved after mutation. The k_cat values of F282L/F306L and Q275P for the substrate ABTS were 574.00 s^-1 and 898.03 s^-1, respectively, which were 1.1 times and 1.7 times that of the wild-type laccase, indicating the improved catalytic efficiency. Q275P demonstrated better performance than the wild-type laccase in pulping, as manifested by the reduction of 0.82 in the Kappa number and the increases of 2.00% ISO, 7.8%, and 7.2% in whiteness, tensile index, and breaking length, respectively. This work lays a foundation for improving the adaptation of laccase to the environment of the papermaking industry.
Laccase/chemistry* ; Directed Molecular Evolution ; Enzyme Stability ; Bacillus pumilus/genetics* ; Mutation ; Biocatalysis ; Catalysis

Functional dyspepsia(FD)is a common functional gastrointestinal disease in clinical practice,which has the characteristics of high incidence,difficult to cure,and recurrence.FD belongs to the categories of"ruffian"and"stomach pain"in TCM,and the disease is located in the stomach,which is closely related to the liver and spleen,and the syndrome of liver depression and spleen deficiency is the most common.This article summarized the literature related to the TCM treatment for FD with liver depression and spleen deficiency syndrome,and concluded the clinical application,efficacy characteristics and mechanism,so as to provide reference for clinical treatment and basic research.The analysis found that the clinical efficacy of TCM in the treatment of FD is significant,which can not only improve the digestive symptoms of patients,but also improve their anxiety and depression state and daily life quality,and has the characteristics of overall regulation,syndrome differentiation and treatment,and improvement of physical fitness.Its mechanism may involve multiple pathways and levels such as abnormal gastric motility,abnormal brain-intestinal interaction and immune inflammatory response.

Ferritin, a ubiquitous protein in living organisms, plays a crucial role in storing and converting iron, as well as maintaining cellular iron metabolism balance. Due to the ability of self-assembling into unique nanocage-like structures in vitro and the special physicochemical properties, ferritin has garnered extensive attention in the biomedical field. This paper provides a brief overview of the structure and cargo loading strategies of ferritin, with a specific focus on its applications in various biological fields such as nanomedicine, bioimaging, and nanoparticle vaccine carriers. The aim is to offer a valuable reference for the future research involving ferritin nanoparticles.
Ferritins/chemistry* ; Nanoparticles/chemistry* ; Humans ; Nanomedicine/methods* ; Animals

Objective To study the current status of the randomized controlled trial(RCT)outcome indicators of acupuncture and moxibustion in the treatment of chronic fatigue syndrome(CFS);To provide guidance and references for clinical research and protocol design.Methods RCT of acupuncture and moxibustion for CFS were retrieved from CNKI,Wanfang Data,VIP,SinoMed,PubMed,Embase,Web of Science databases from January 1,2015 to December 9,2024.Through literature screening and data extraction,Excel 2019 was used to make a descriptive analysis of the basic characteristics,diagnostic criteria,intervention measures,classification of outcome indicators,use frequency and evaluation time points of the included studies.Origin2021 software was used to draw bubble charts for visual display.Results Totally 119 RCT were included,involving 8 445 subjects,5 diagnostic criteria of Western medicine,5 diagnostic criteria of TCM,and 6 outcome indicators,namely clinical symptoms/signs,symptoms/syndrome of TCM,physical and chemical examination,quality of life,safety evaluation,and patient satisfaction.A total of 58 outcome indicators were reported,with a cumulative frequency of 430 times.Conclusion The RCT of acupuncture and moxibustion treatment of CFS have some problems,such as incomplete baseline assessment,lack of sample size calculation basis,high risk of bias,ignorance of primary and secondary indicators in the selection of outcome indicators,numerous indicators selection and measurement points,lack of long-term follow-up,inconsistent evaluation criteria for TCM syndrome efficacy,and insufficient attention to health economics evaluation and safety outcome indicators.

Functional dyspepsia(FD)is a common functional gastrointestinal disease in clinical practice,which has the characteristics of high incidence,difficult to cure,and recurrence.FD belongs to the categories of"ruffian"and"stomach pain"in TCM,and the disease is located in the stomach,which is closely related to the liver and spleen,and the syndrome of liver depression and spleen deficiency is the most common.This article summarized the literature related to the TCM treatment for FD with liver depression and spleen deficiency syndrome,and concluded the clinical application,efficacy characteristics and mechanism,so as to provide reference for clinical treatment and basic research.The analysis found that the clinical efficacy of TCM in the treatment of FD is significant,which can not only improve the digestive symptoms of patients,but also improve their anxiety and depression state and daily life quality,and has the characteristics of overall regulation,syndrome differentiation and treatment,and improvement of physical fitness.Its mechanism may involve multiple pathways and levels such as abnormal gastric motility,abnormal brain-intestinal interaction and immune inflammatory response.

Objective:To analyze identification of copper death gene related subtypes,construction of prognosis model and influence of immune infiltration in osteosarcoma(OS)on basis of copper death gene.Methods:Survival and prognosis of OS associated copper death gene were analyzed combining by TARGET and GEO database.OS was divided into different subtypes of copper death by consistent clustering method.SSGSEA was used to analyze difference of immune cells in classification of copper death.Setting P value= 0.05 and q value=0.05,GO and KEGG enrichment analysis were performed on differential genes of copper death typing.Prognosis model was constructed according to results of Lasso regression analysis and cross validation,risk assessment analysis and ROC curve were used to evaluate accuracy of model prediction.Combined with clinical characteristics,nomograms were constructed to predict survival time of patients,and risk differences were analyzed.Immune cell infiltration and tumor microenvironment analysis were performed on OS samples."pRRophetic"package in R software was used to analyze drug sensitivity of OS samples.Results:FDX1,GLS,DLAT and PDHB as high-risk genes for OS prognosis were identified.According to copper death classification of OS samples,OS could be divided into two types:CRGclusterA and CRGclusterB.CRGclusterA was associated with Th2 cells,and CRGclusterB was associated with Th1 cells.Most OS copper death genes were highly expressed in CRGclusterA.Immune cell infiltration analysis results showed that γδ T cells,resting mast cells and resting dendritic cells were positively correlated with risk score,while CD8 T cells were negatively correlated with risk score.Drug sensitivity analysis showed that OS showed higher sensitivity to Elesclomol and GW.441756.Conclusion:Two subtypes of CRGclusterA and CRGclusterB are identified in this study.Four high-risk prognostic genes FDX1,GLS,DLAT and PDHB are identified,providing new insights into prognostic evaluation and immunotherapy target candidates for OS.

Objective:To study the differential gene expression and immune cell infiltration of gout patients,to find the key genes and immune cells of gout pathogenesis,and to explore the relationship between immune cells and gout.Methods:The gout chip GSE160170 was downloaded from the GEO database,and the differential gene expression analysis was carried out with the help of R language.Then,the STRING database was used to analyze the differential gene,and the Cytoscape software was used to screen the key genes,and then carry out enrichment analysis.At the same time,the infiltration of immune cells were analyzed.Results:The study found that IL-6,IL-1β,TNF,CCL3,CXCL8 and CXCL1 were key genes in the pathogenesis of gout,which were mainly exerted by IL-17,Toll-like receptor,NOD-like receptor,NF-κB and other signaling pathways.Processes such as cellular responses to lipo-polysaccharides,bacteria-derived molecules,and biological stimuli lead to disease;immune infiltration results indicate that memory B cells,activated NK cells,activated dendritic cells,activated mast cells and eosinophils were involved in the disease.It was signifi-cantly expressed in gout patients;the correlation analysis between immune cells showed that the expression of follicular helper T cells were positively correlated with the expression of activated mast cells,and the expression of unactivated NK cells and monocyte were negatively correlated.Conclusion:Key genes and differentially expressed immune cells are closely related to the pathogenesis of gout,providing new ideas for the study of the molecular mechanism of gout.

ObjectiveTo analyze the traditional Chinese medicine （TCM） patterns in patients with spasmodic torticollis and provide reference for standardized differentiation and clinical treatment. MethodsA cross-sectional study was conducted in the spasmodic torticollis outpatient clinic and dystonia ward of the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine from June 2022 to December 2023. The general information including gender， age， duration of disease and type， and the TCM four examinations data such as symptoms， tongue manifestation and pulse manifestation of 198 patients with spasmodic torticollis were obtained by means of on-site questionnaires. Descriptive frequency analysis， factor analysis， and cluster analysis were performed， and the distribution of major TCM patterns were summarized based on the clinical information. ResultsA total of 198 patients with spasmodic torticollis were included， of which 89 （44.95%） were male and 109 （55.05%） were female， with an average age of 40.70±0.96 years and an average course of disease of 24.78±2.32 months. A total of 296 symptoms/signs were obtained， with a cumulative frequency of 6756 times， of which 58 symptoms/signs had a frequency ≥20%， and the top three were neck and back stiffness （83.84%）， condition related to emotions （74.75%） and irritability （72.73%）. Factor analysis of 58 symptoms/signs showed that factor rotation converged after 51 iterations， resulting in 20 common factors with a cumulative contribution of 64.03%. The top three syndrome elements related to the location of the disease were liver， channels， tendons and bones， and those related nature of the disease were dominated by qi stagnation， blood stasis and yin deficiency. The cluster analysis of the 20 common factors showed that the main TCM patterns were internal stirring of liver wind syndrome， liver-kidney yin deficiency syndrome， turbid phlegm obstruction syndrome， and pathogen congested in the channels syndrome. Among the 198 patients， 81 were diagnosed with internal stirring of liver wind syndrome， 60 with liver-kidney yin deficiency syndrome， 37 with turbid phlegm obstruction syndrome， and 20 with pathogen congested in the channels syndrome.There was no statistically significant difference in the distribution of TCM patterns among patients of different genders， age groups， and duration of disease （P＞0.05）. ConclusionSpasmodic torticollis is mainly located in the liver， mostly with internal stirring of liver wind syndrome， liver-kidney yin deficiency syndrome， turbid phlegm obstruction syndrome， and pathogen congested in the channels syndrome.