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.

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

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

BACKGROUND:The pathogenesis of osteoporosis is complex,and its essence is the weakening of bone formation and the enhancement of bone absorption caused by various reasons,resulting in the imbalance of bone metabolism.In recent years,N6-methyladenosine has been found(N6-methyladenosine,m6A)methylation can prevent and treat osteoporosis by regulating bone metabolism. OBJECTIVE:Taking the regulation of bone metabolism by m6A methylation as an entry point,to systematically sort out and summarize the research progress of m6A methylation in osteoporosis,so as to provide certain theoretical reference bases for the search of new therapeutic targets for osteoporosis. METHODS:CNKI,WanFang,VIP,PubMed,MEDLINE,Nature,and Cochrane databases were retrieved for relevant literature published from database inception to 2023.The keywords were"osteoporosis,m6A methylation,bone metabolism,bone marrow mesenchymal stem cells,osteoblasts,osteoclasts"in Chinese and English.Duplicates and obsolete non-referenced documents were excluded,and a total of 73 standard papers were included for further review. RESULTS AND CONCLUSION:m6A methylation can affect the activity and differentiation of bone marrow mesenchymal stem cells,osteoblasts,and osteoclasts through various pathways to regulate bone metabolism and prevent osteoporosis.The regulatory process of m6A methylation is extremely complex,and its related proteins play different roles in different cells.Even in the same kind of cells,the same type of proteins may have radically different roles,regulating different physiological and pathological processes.

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.

Objective To investigate the mechanism of abdominal massage for treating chronic fatigue syndrome(CFS).Methods Thirty clean-grade female Sprague-Dawley rats were randomly divided into the normal control,model control,and abdominal massage groups(n=10 rats per group).The CFS rat model was established through cold water swimming combined with the chronic restraint method in the model control and abdominal massage groups.The rats in the abdominal massage group were treated with the core techniques of Jingu zang-fu massage,namely layer press"Guanyuan"(CV4)for 8 min and Tuanmo"Zhongwan"(CV12)for 12 min as the primary intervention techniques,once a day for 14 consecutive days.The rats in the two control groups did not receive intervention;however,they were bound to the experimental bench when the experimental group was massaged.After the intervention,the indexes of the main organs of the hypothalamic-pituitary-adrenal(HPA)axis and hippocampal cell apoptosis in each group were measured.The morphology of hippocampal cells in each group was observed using Nissl staining of hippocampal tissue and transmission electron microscopy of hippocampal neurons.Results The index of each organ in the model control group was upregulated(P<0.01)compared to that of the normal control group.In contrast,the index of each organ in the abdominal massage group was downregulated(P<0.01)compared with that of the model control group.Compared to the normal control group,the index of each organ in the abdominal massage group was upregulated;however,the difference was not significant.Compared to the normal control group,the cell nuclei in the model control group were significantly consolidated,the nuclear membrane structure was ruptured,and the margins were irregular.Most of the cell morphology in the abdominal massage group was normal compared with that of the model control group,and a small number of nuclear membrane structures were unclear.Compared to the normal control group,the hippocampal neuronal cell in the other two groups was significantly damaged,and the hippocampal neuronal cell in the abdominal massage group was in good condition compared to the model control group.The degree of neuronal apoptosis in the model control group was significantly higher than that in the normal control group(P<0.01).The degree of neuronal apoptosis in the abdominal massage group decreased compared to that of the model control group(P<0.01),which was slightly higher than that in the normal control group but not significant.Conclusion Cold water swimming combined with chronic restraint can simulate CFS in rats,and abdominal massage can reduce the organ index of the hypothalamus,pituitary gland,and adrenal gland,increase hippocampal cell activity,reduce hippocampal tissue damage,inhibit hippocampal cell apoptosis,and maintain the normal physiological function of hippocampal neurons.

Objective:To study the common pathogenesis of gout and rheumatoid arthritis(RA)by bioinformatics analysis.Methods:Microarray expression profiles of peripheral blood mononuclear cells in gout and RA were obtained from the GEO public da-tabase.R language and other tools were used to re-annotates the chip,and then the differential genes(DEGs)of the two were screened and the intersection was taken.The protein-protein interaction(PPI)network and topology analysis of common differential genes(CO-DEGs)were constructed by STRING database and Cytoscape software(including CytoNCA plug-in).The HubGene was screened and validated by ROC curve.Finally,the DAVID online analysis tool was used to perform GO and KEGG functional enrichment analysis of HubGene.Results:There were 9 HubGene screened,they were TNF,RGS1,CD69,IL7R,DDX3X,SOCS3,IFIT1,IFIT3,CCL3.GO enrichment showed that HubGene was mainly involves the regulation of virus,STAT receptor signaling pathway and positive regu-lation of neuroinflammatory response.KEGG enrichment showed that HubGene was mainly involved in Toll like receptor signaling pathway,TNF signaling pathway,JAK-STAT signaling pathway,adipocytokine signaling pathway,RIG-Ⅰ-like receptor signaling pathway and osteoclast differentiation.Conclusion:Using bioinformatics analysis,nine HubGene and related signaling pathways in-volved in the pathogenesis of gout and RA have been identified,which may serve as novel biomarkers and potential targets.

Objective:To analyze the acoustic characteristics of patients with hypopharyngeal cancer accompanied by vocal cord dysfunction. Methods:A retrospective analysis was conducted on the data of patients with hypopharyngeal cancer who were initially treated at The Second Affiliated Hospital of Xi'an Jiaotong University from January 2018 to April 2024. Patients who had completed electronic laryngoscopy, stroboscopic laryngoscopy, and voice analysis were selected from the data. Among them, patients with hypopharyngeal cancer who had unilateral vocal cord activity disorders were selected as the experimental group, while patients with symmetrical bilateral vocal cord activity were assigned to the control group. Then the clinical characteristics, the vocal parameters, and the stroboscopic laryngoscopy results of patients with hypopharyngeal cancer in the experimental group and the control group were analyzed and compared. Results:Compared with that in the control group, the proportion of lesions located on the inner wall of the piriform fossa in the experimental group increased（83.3% vs 53.8%）, and the difference was statistically significant（P<0.05）. There was no significant difference in vocal parameters such as SPL, Jitter, Shimmer, MPT, DSI, F0, sound intensity, electroglottic value and VHI between the experimental group and the control group（P>0.05）. However, the values of F0, Jitter, Shimmer and VHI in the experimental group were higher than those in the control group. In addition, in terms of the results of stroboscopic laryngoscopy, the proportion of glottic insufficiency（42.9% vs 18.8%） and asymmetric arytenoid cartilage（64.3% vs 0） in the experimental group was significantly higher than that in the control group（P<0.05）. However, the mucosal waves of the vocal cords on the affected side did not weaken in patients in both the experimental group and the control group. In the experimental group of 18 patients with hypopharyngeal cancer who received induction chemotherapy（nituzumab+nedaplatin+5-fluorouracil）, 13 of them had improved vocal cord activity（improvement rate of 72.2%）. Conclusion:Hypopharyngeal cancer in the medial wall of the pyriform fossa is more prone to vocal cord dysfunction, but vocal cord dysfunction has little effect on the vocal parameters of patients with hypopharyngeal cancer.
Humans ; Hypopharyngeal Neoplasms ; Retrospective Studies ; Male ; Female ; Laryngoscopy ; Middle Aged ; Vocal Cord Dysfunction/etiology* ; Vocal Cords/physiopathology* ; Stroboscopy ; Voice Quality ; Aged