Objective To assess the therapeutic efficacy of low-intensity focused ultrasound(LIFU)on knee arthrofibrosis in rats and explore its underlying mechanisms.Methods Fifteen male SD rats(8 weeks old,weighing 250～300 g)were randomly divided into a blank control group,a model group,and an ultrasound treatment group,with 5 animals in each group.Rat model of knee joint stiffness was established in the model and ultrasound treatment group.The rats in the ultrasound group received LIFU intervention(frequency:1 MHz,power:1.5 W,20 min per session,5 times/week)for 4 weeks.The knee ranges of motion(ROM,flexion and extension)were measured in all 3 groups at 0,14 and 28 d after intervention.Histological analysis was conducted for the morphological changes in the posterior capsular area of the knee.After the synovial fibroblasts were primary isolated from rat knee joints,the cells were divided into blank control,LIFU,TGF-β(10 ng/mL),and TGF-β+LIFU groups.Changes in fibrosis-related indicators and the TGF-β/Smad signaling pathway were assessed in each group.Results In the animal experiments,LIFU intervention for 14 d resulted in significantly improved flexion-extension ROM in knee joints when compared with the rats in the model group(P<0.05),and the improvement was more obvious at 28 d after intervention(P<0.05).After 28 d of LIFU intervention,the fibrosis of the posterior capsule of the knee joint was notably improved in the ultrasound group than the model group,and the expression levels of fibrosis-related indicators(α-SMA,TGF-β)were decreased(P<0.05).In the cellular experiments,the TGF-β group exhibited remarkable up-regulation of fibrosis-related molecules(α-SMA,COL1A1,COL3A1)at both protein and mRNA levels(P<0.05),and activation of the TGF-β/Smad signaling pathway when compared to the blank control group.While LIFU intervention inhibited the expression of TGF-β-induced upregulation of fibrotic indicators(α-SMA,COL1A1,COL3A1)and the activation of TGF-β/Smad signaling pathway(P<0.05).Conclusion LIFU can effectively improve knee arthrofibrosis in rats,which potentially through the inhibition of fibroblast activation and modulation of the TGF-β/Smad signaling pathway.

Sonodynamic therapy (SDT) can potentially induce immunogenic cell death in tumor cells, leading to the release of ATP, and facilitating the initiation of an immune response. Nevertheless, the enzymes CD39 and CD73 can swiftly convert ATP into immunosuppressive adenosine (ADO), resulting in an immunosuppressive tumor microenvironment (TME). This study introduced a nanomedicine (QD/POM1@NP@M) engineered to reprogram TME by modulating the CD39/CD73/ADO pathway. The nanomedicine encapsulated sonosensitizers silver sulfide quantum dots, and the CD39 inhibitor POM1, while also incorporating homologous tumor cell membranes to enhance targeting capabilities. This integrated approach, on the one hand, stimulates the release of ATP via SDT, thereby initiating the immune response. In addition, it reduced the accumulation of ADO by inhibiting CD39 activity, which ameliorated the immunosuppressive TME. Upon administration, the nanomedicine demonstrated substantial anti-tumor efficacy by facilitating the infiltration of anti-tumor immune cells, while reducing the immunosuppressive cells. This modulation effectively transformed the TME from an immunologically "cold" state to a "hot" state. Furthermore, combined with the checkpoint inhibitor α-PDL1, the nanomedicine augmented systemic anti-tumor immunity and promoted the establishment of long-term immune memory. This study provides an innovative strategy for combining non-invasive SDT and ATP-driven immunotherapy, offering new ideas for future cancer treatment.

Primary immune thrombocytopenia(ITP)is an acquired autoimmune hemorrhagic disease characterized by reduced isolated peripheral blood platelet counts for which pathogenesis and etiology are not well defined.Present diagnosis of ITP is to be excluding other diseases,whose pathogenesis generally involves multiple aspects of immunology,biology,epigenetics and environ-mental factors.Recognition of epigenetic alterations in dysregulation of immune tolerance and autoimmune diseases has been deepened in recent years with progress by molecular biology.DNA methylation,an important epigenetic modification,has been shown to play a role in pathogenesis of ITP.Abnormal DNA methylation leads to cellular abnormalities in proliferation,differentiation,apoptosis and immune response in body,induces immune intolerance to autologous platelet antigens in patients,resulting in abnormal activation of immune cells which makes excessive platelet destruction or insufficient platelet production.Therefore,this article reviews latest research progress of DNA methylation genes in ITP to provide a reference for further recovery of immune tolerance of patients.

AIM:To investigate the effects of macrophage galactose-type lectin 1(Mgl1)gene deletion on he-matopoietic stem/progenitor cells(HSPCs)under steady-state conditions and inflammation.METHODS:Mice were di-vided into a control group(wild-type)and an experimental group(Mgl1 gene-deleted).Flow cytometry was used to ana-lyze the proportions of various hematopoietic cell lineages in the peripheral blood and bone marrow of both groups,assess-ing the impact of Mgl1 gene deletion on steady-state hematopoiesis(n=3～4).Transplantation and colony-forming assays were utilized to evaluate the effects of Mgl1 gene deletion onthe repopulation capacity and colony-forming ability of HSPCs(n=5).The LPS-induced inflammation model was employed to examine the effects of Mgl1 gene deletion on the inflamma-tory response of HSPCs both in vitro and in vivo(n=5～8).Western blot and RT-qPCR were conducted to analyze the alter-ations in signaling pathways regulated by Mgl1 in the inflammatory response of HSPCs(n=3).RESULTS:(1)Mgl1 gene deletion had no significant effecton steady-state hematopoiesis(P>0.05).(2)Mgl1 gene deletion promoted inflam-mation-induced cell differentiation of HSPCs(P<0.01).(3)Mgl1 gene deletion accelerated the exhaustion of HSPCs un-der prolonged inflammatory conditions(P<0.01).(4)Mgl1 was found to regulate the inflammatory response of HSPCs via the NF-κB signaling pathway.CONCLUSION:Mgl1 gene deletion enhances the inflammatory response of HSPCs via the NF-κB signaling pathway.

Primary immune thrombocytopenia(ITP)is an acquired autoimmune hemorrhagic disease characterized by reduced isolated peripheral blood platelet counts for which pathogenesis and etiology are not well defined.Present diagnosis of ITP is to be excluding other diseases,whose pathogenesis generally involves multiple aspects of immunology,biology,epigenetics and environ-mental factors.Recognition of epigenetic alterations in dysregulation of immune tolerance and autoimmune diseases has been deepened in recent years with progress by molecular biology.DNA methylation,an important epigenetic modification,has been shown to play a role in pathogenesis of ITP.Abnormal DNA methylation leads to cellular abnormalities in proliferation,differentiation,apoptosis and immune response in body,induces immune intolerance to autologous platelet antigens in patients,resulting in abnormal activation of immune cells which makes excessive platelet destruction or insufficient platelet production.Therefore,this article reviews latest research progress of DNA methylation genes in ITP to provide a reference for further recovery of immune tolerance of patients.

AIM:To investigate the effects of macrophage galactose-type lectin 1(Mgl1)gene deletion on he-matopoietic stem/progenitor cells(HSPCs)under steady-state conditions and inflammation.METHODS:Mice were di-vided into a control group(wild-type)and an experimental group(Mgl1 gene-deleted).Flow cytometry was used to ana-lyze the proportions of various hematopoietic cell lineages in the peripheral blood and bone marrow of both groups,assess-ing the impact of Mgl1 gene deletion on steady-state hematopoiesis(n=3～4).Transplantation and colony-forming assays were utilized to evaluate the effects of Mgl1 gene deletion onthe repopulation capacity and colony-forming ability of HSPCs(n=5).The LPS-induced inflammation model was employed to examine the effects of Mgl1 gene deletion on the inflamma-tory response of HSPCs both in vitro and in vivo(n=5～8).Western blot and RT-qPCR were conducted to analyze the alter-ations in signaling pathways regulated by Mgl1 in the inflammatory response of HSPCs(n=3).RESULTS:(1)Mgl1 gene deletion had no significant effecton steady-state hematopoiesis(P>0.05).(2)Mgl1 gene deletion promoted inflam-mation-induced cell differentiation of HSPCs(P<0.01).(3)Mgl1 gene deletion accelerated the exhaustion of HSPCs un-der prolonged inflammatory conditions(P<0.01).(4)Mgl1 was found to regulate the inflammatory response of HSPCs via the NF-κB signaling pathway.CONCLUSION:Mgl1 gene deletion enhances the inflammatory response of HSPCs via the NF-κB signaling pathway.

ObjectiveTo assess the level of internal exposure to PCP in a community population in Shanghai， to investigate the factors affecting the level of PCP， and to analyze the correlation between the exposure and thyroid hormone levels. MethodsA total of 464 residents of a community in Shanghai were selected as the study subjects. A questionnaire survey was conducted to obtain the demographic information， dietary situation， lifestyle and behavioral habits， and disease history of the individuals， and blood samples were collected. Gas chromatography-electron trap was applied to determine the PCP levels in serum. Multicategorical logistic regression analysis was used to analyze the possible influencing factors of PCP exposure in humans. Thyroid hormone levels were used as the dependent variable and serum PCP as the independent variable. Multiple linear regression analysis was used to assess the association between PCP and thyroid hormones in the community population after controlling the confounding factors such as age， gender， literacy， annual personal income， and chronic diseases. ResultsThe detection rate of serum PCP in 464 subjects was 90.3%， and the median serum PCP level was 0.43 μg·L^-1. The differences in PCP levels among different age groups were statistically significant. There were no significant differences in PCP levels among different gender and BMI groups. The study of PCP exposure factors showed that age， frequency of using plastic products， consumption of freshwater fish， type of occupation， annual income， and consumption of tea or coffee were the potential influencing factors for PCP exposure. Among them， age， frequency of using plastic products， consumption of tea or coffee， and consumption of freshwater fish were positively associated with PCP levels， and annual personal income was negatively associated with it. The results of multiple linear regression analysis showed that among men， PCP levels were positively correlated with TSH （b=0.105， 95%CI：0.017‒0.313） and negatively correlated with FT4 （b=-0.026， 95%CI：-0.057‒0.004）， and among women， PCP levels were positively correlated with TSH （b=0.092， 95%CI：-0.211‒0.904） and FT3 （b=0.017， 95%CI：-0.058‒0.230） and negatively correlated with FT4 （b=-0.013， 95%CI：-0.011‒0.037）. ConclusionSerum PCP detection is common among community residents in Shanghai. Different demographic characteristics or behavioral habits may increase or decrease PCP exposure. PCP exposure then affects human thyroid hormone levels.

Objective:To explore the distribution pattern of TCM syndromes in patients with pulmonary tuberculosis combined with coronary heart disease in Kunming area.Methods:A survey was conducted by research of questionnaires to the general information and TCM four diagnostic information of pulmonary tuberculosis patients (116 cases) with coronary heart disease admitted to our hospital from April 2019 to December 2020, and principal component analysis and clustering analysis were conducted. Frequency analysis and severity analysis methods were used for data processing.Results:The TCM symptoms of pulmonary tuberculosis patients with coronary heart disease were mainly cough [89.66%(104/116)], expectoration [73.28% (85/116)], dry mouth [70.69% (82/116)], chest tightness [64.66% (75/116)], fatigue [56.03%(65/116)], hot flashes [52.59% (61/116)], shortness of breath [50.00% (58/116)]; tongue color was more common with light red [43.10% (50/116)] and red [47.41% (55/116)]; the tongue shape was more common with cracked tongue [37.07% (43/116)], punctured tongue [27.59% (32/116)] and old tongue [23.28%(27/116)]; tongue coating with little or no coating [48.28%(56/116)], yellow and greasy coating [21.55% (25/116)] was more common; the pulse was usually thin (number) [54.31% (63/116)] and slippery (number) [25.86% (30/116)]. The principal component analysis method used a load coefficient >0.40 as the threshold to screen out the four diagnostic information of 10 principal components; the clustering analysis results were divided into three categories of TCM types: deficiency syndrome, excess syndrome, and mixed deficiency and excess syndrome. There were significant differences in TCM syndrome types among patients of different genders, ages, disease courses, and educational levels ( P<0.05). Deficiency syndrome was more common in male patients [41.18% (21/51)], and mixed syndrome of deficiency and excess was more common in female patients [63.08% (41/65)]; 43-59 years old patients were more likely to have excess syndrome [36.36%(24/66)], and ≥60 years old patients were more likely to have mixed syndrome [70.00% (35/50)]; patients with a course of disease <3 months had more excess syndrome [41.30% (19/46)], patients with a course of 3-12 months had more deficiency syndrome [57.14% (20/35)], and patients with a course of more than 1 year had more mixed syndrome of deficiency and excess [74.29% (26/35)]. male patients [54.17%(26/48)] were more serious in excess syndrome, and female patients [53.33% (24/45)]; were more serious in deficiency syndrome; deficiency syndrome [43-59 years old was 54.17% (42/84), ≥60 years old was 54.17% (12/24)] more serious in patients of different age groups; patients with a course of disease <3 months [56.86% (29/51)] were more serious in excess syndrome, and patients with a course of disease ≥3 months were more serious in mixed syndromes [3~12 months 52.38%(22/42), >1 year 53.33% (24/45)]. Conclusion:The pathogenesis of TCM in patients with pulmonary tuberculosis complicated with coronary heart disease in Kunming is deficiency in root and excess in superficiality; the syndrome is based on yin deficiency and qi deficiency, with blood stasis, phlegm heat, phlegm stasis and phlegm turbidity as the symptoms.

AIM:To investigate the effects of sodium selenite(SS)on viability,migration and angiogenesis of human non-small-cell lung cancer(NSCLC)H520 and A549 cells.METHODS:The H520 cells,A549 cells,and hu-man umbilical vein endothelial cells(HUVEC)were divided into control group(0 μmol/L SS),low dose group(5 μmol/L SS),middle dose group(10 μmol/L SS),and high dose group(20 μmol/L SS).Cell viability was assessed using the CCK-8 assay,and the half-maximal inhibitory concentration(IC50)was calculated.Cell migration and invasion abilities were determined through wound healing and Transwell assays.The regulatory effects of SS on angiogenesis,vasculogenic mimicry and"mosaic"vascular formation between HUVEC and NSCLC cells were detected using vessel forming assays.The expression of vascular endothelial growth factor(VEGF)in the supernatant of lung cancer cells in each group was de-tected using chemiluminescence.RT-qPCR was used to assess the mRNA expression of VEGF,vascular endothelial growth factor receptor 2(VEGFR2)and angiotensin II(Ang II).Western blot was used to examine the protein levels of VEGF,p-PI3K,and p-Akt in H520 and A549 cells.RESULTS:The IC50 values of SS to HUVEC,A549 cells and H520 cells for 48 h were 6.762,9.003 and 7.356 μmol/L,respectively.Compared with control group,the wound healing rate was significantly decreased in each group treated with SS for 48 h(P<0.01).In HUVEC,the number of migrating cells in middle dose and high dose groups decreased(P<0.01),whereas in lung cancer cells,the number of migrating cells in each group decreased after SS treatment(P<0.01).The mRNA expression levels of VEGF,VEGFR2 and Ang II were lower in high-dose SS group than those in control group(P<0.05 or P<0.01).In H520 cells,compared with control group,the protein levels of VEGF,p-PI3K and p-Akt in SS treatment groups were significantly decreased(P<0.05 or P<0.01).CONCLUSION:Sodium selenite inhibits the viability and migration of HUVEC,H520 cells and A549 cells,and inhibits the formation of vasculogenic mimicry and mosaic vessels in NSCLC cells.This mechanism may be related to the inhibition of PI3K-Akt signaling pathway activation and regulation of VEGF.

Inducing the degradation of KRAS represents a novel strategy to combat cancers with KRAS mutation. In this study, we identify ubiquitin-specific protease 2 (USP2) as a novel deubiquitinating enzyme of KRAS in multiple myeloma (MM). Specifically, we demonstrate that gambogic acid (GA) forms a covalent bond with the cysteine 284 residue of USP2 through an allosteric pocket, inhibiting its deubiquitinating activity. Inactivation or knockdown of USP2 leads to the degradation of KRAS, resulting in the suppression of MM cell proliferation in vitro and in vivo. Conversely, overexpressing USP2 stabilizes KRAS and partially abrogates GA-induced apoptosis in MM cells. Furthermore, elevated USP2 levels may be associated with poorer prognoses in MM patients. These findings highlight the potential of the USP2/KRAS axis as a therapeutic target in MM, suggesting that strategically inducing KRAS degradation via USP2 inhibition could be a promising approach for treating cancers with KRAS mutations.