Objective:To explore the potential mechanism underlying IL-6 production through the TLR7 signaling pathway,which regulates Th17 cell differentiation in the context of Coxsackievirus B3(CVB3)-induced acute viral myocarditis(AVMC).Meth-ods:A total of 110 patients diagnosed with AVMC were admitted to Quanzhou First Hospital,Fujian between January 2020 and Janu-ary 2023,alongside 93 healthy volunteers.CD4+T cells were isolated from the subjects'blood,and the levels of CVB3 and the number of Th17 cells were assessed.Subsequently,CD4+T cells were infected with CVB3,and the levels of Th17 cells,IL-17,IL-21,and TNF-α were measured.After knockdown of TLR7 or treatment with TLR7 inhibitors,the differentiation of CVB3-infected CD4+T cells into Th17 cells was observed.Results:In comparison to healthy controls,AVMC patients exhibited elevated plasma levels of hsCRP,IL-17,IL-21,and TNF-α(P<0.05).The levels of CVB3 mRNA in CD4+T cells were also notably higher in AVMC patients compared to healthy controls(P<0.05).The mean viral titer in AVMC patients measured 230 PFU/ml,while no detectable virus was found in healthy volunteers(P<0.05).In CD4+T cells,the count of Th17 cells was significantly increased in AVMC patients compared to healthy volunteers(P<0.05).Moreover,the number of Th17 cells in peripheral blood CD4+T cells of AVMC patients showed a positive correlation with CVB3 virus titer(P<0.05).Following CVB3 infection,the number of Th17 cells increased compared with the control group(P<0.05),accompanied by elevated levels of IL-17,IL-21,and TNF-α in the supernatant(P<0.05).Knockdown of TLR7 and CVB3 infection in CD4+T cells significantly reduced the levels of Th17 cells(P<0.05),while the expression level of phosphorylated-activated TLR7 increased significantly after CVB3 infection of CD4+T cells compared to the control group(P<0.05).Treatment with the TLR7 inhibitor M5049 and CVB3 infection led to a significant decrease in Th17 cell levels(P<0.05).The secretion of IL-6 in CD4+T cells increased after CVB3 infection(P<0.05),and this increase was mitigated by TLR7 knockdown and CVB3 infection(P<0.05)as well as TLR7 inhibitor M5049 treatment and CVB3 infection(P<0.05).Conclusion:CVB3 activates TLR7 via phosphoryla-tion,prompting CD4+T cells to release IL-6 and undergo differentiation into Th17 cells.Consequently,TLR7 emerges as a promising therapeutic target for AVMC.

Blood-bi-syndrome is primarily characterized by localized numbness and mild pain in the limbs.Records about the diagnosis and treatment of blood-bi-syndrome are scattered throughout classical medical literature.According to the theory of Yindan and Yangdan formulas in the classical prescription system,when the body's ascending function of qi movement is insufficiency,Yangdan formulas which contain Astragali Radix can be used for warming and uplifting yang qi.Conversely,when the descending function of qi movement is insufficiency,Yindan formulas which contain containing Bupleuri Radix can be used for astringing,descending,clearing and purging.This article,based on the principles of Yindan and Yangdan formulas,analyzed the fundamental pathogenesis of blood-bi-syndrome developing from the transmission of superficial syndromes,as well as its clinical manifestations during the transmission and change between taiyin disease and yangming disease.And the therapeutic strategies for blood-bi-syndrome were also summarized.It is proposed that taiyin blood-bi-syndrome arises in individuals with constitutionally deficient cold after the attack of pathogenic factors,resulting from fluid depletion,blood insufficiency,and stomach deficiency,which should be treated by Yangdan Formulas with pungent-sweet-warm properties for warming and uplifting yang qi.For the treatment of patients with taiyin blood-bi-syndrome characterized by exterior syndrome,Astragali Radix-containing classical prescriptions like Astragali Radix and Cinnamomi Ramulus Five-Component Decoction(Huangqi Guizhi Wuwu Decoction)can be chosen;for the treatment of patients with taiyin blood-bi-syndrome characterized by interior syndrome,Zingiberis Rhizoma Recens-containing classical prescriptions like Citri Reticulatae Pericarpium and Bambusae Caulis in Taenia Decoction(Jupi Zhuru Decoction)is adopted.Conversely,yangming blood-bi-syndrome occurs in individuals with constitutionally excessive heat after the attack of pathogenic factors,resulting from the internal accumulation of blood stasis and heat and the impairment of the fluid and blood,which should be treated by Yindan Formulas with pungent-sweet-cold or sour-cold properties for astringing,descending,clearing and purging.For the treatment of patients with yangming blood-bi-syndrome characterized by exterior syndrome,Puerariae Lobatae Radix-containing classical prescriptions like Phyllostachydis Henonis Folium and Puerariae Lobatae Radix Decoction(Zhuye Gegen Decoction)can be used;for the treatment of patients with yangming blood-bi-syndrome characterized by interior syndrome,Paeoniae Radix Alba-containing classical prescriptions like Rhei Radix et Rhizoma and Eupolyphaga seu Steleophaga Decoction(Dahuang Zhechong Wan)is recommended.

Objective To investigate the effects of TEK receptor tyrosine kinase(Tie2)L914F mutation on the biological behavior of vascular endothelial cells and the changes of related signaling pathways in patients with venous malformations.Methods Gene sequen-cing was used to detect Tie2-L914F mutations in venous malformations.HE staining and immunohistochemical staining were used to de-tect the expression of platelet derived growth factor subunit B gene(PDGFB)and α-smooth muscle actin(α-SMA)in venous malfor-mations caused by the mutation.Lentivirus overexpressing Tie2-wild type(Tie2-WT),Tie2-L914F and Tie2-GFP(green fluorescent protein,GFP)infected human umbilical vein endothelial cells(HUVECs).Real-time fluorescence quantitative PCR was used to detect the expression level of Tie2 in endothelial cells expressing exogenous Tie2 and Flag-tagged protein Tie2 protein was detected by western blotting to verify transfection efficiency.The proliferation,apoptosis,migration and tube-forming ability of the cells were determined by CCK-8,flow cytometry,Transwell migration assay and Matrigel matrix gel tube-forming assay.Western blotting was used to detect the expression levels of Protein Kinase B(PKB/AKT),FOXO1 and their phosphorylation,and the expression level of PDGFB was detec-ted by ELISA.Results The number of patients with venous malformations with L914F mutations was about 33.3％.HE and immuno-histochemical staining showed that the expressions of PDGFB and α-SMA were significantly down-regulated in venous malformation tis-sues with Tie2-L914F mutation,and were positively correlated with the decrease in cell coverage of the tube wall.Compared with Tie2-WT endothelial cells,the apoptosis number of Tie2-L914F endothelial cells was significantly reduced,while the proliferation and mi-gration ability was significantly increased,and the tube-forming ability was significantly decreased.Western blotting and ELISA showed that the phosphorylation levels of AKT and FOXO1 downstream of Tie2 signaling pathway in endothelial cells expressing Tie2-L914F were significantly increased,and the expression level of PDGFB was significantly decreased.Conclusion In venous malformations,Tie2-L914F mutation may downregulate the expression of PDGFB through AKT signaling pathway,which affects the biological behavior of vascular endothelial cells.

Objective:To explore the potential mechanism underlying IL-6 production through the TLR7 signaling pathway,which regulates Th17 cell differentiation in the context of Coxsackievirus B3(CVB3)-induced acute viral myocarditis(AVMC).Meth-ods:A total of 110 patients diagnosed with AVMC were admitted to Quanzhou First Hospital,Fujian between January 2020 and Janu-ary 2023,alongside 93 healthy volunteers.CD4+T cells were isolated from the subjects'blood,and the levels of CVB3 and the number of Th17 cells were assessed.Subsequently,CD4+T cells were infected with CVB3,and the levels of Th17 cells,IL-17,IL-21,and TNF-α were measured.After knockdown of TLR7 or treatment with TLR7 inhibitors,the differentiation of CVB3-infected CD4+T cells into Th17 cells was observed.Results:In comparison to healthy controls,AVMC patients exhibited elevated plasma levels of hsCRP,IL-17,IL-21,and TNF-α(P<0.05).The levels of CVB3 mRNA in CD4+T cells were also notably higher in AVMC patients compared to healthy controls(P<0.05).The mean viral titer in AVMC patients measured 230 PFU/ml,while no detectable virus was found in healthy volunteers(P<0.05).In CD4+T cells,the count of Th17 cells was significantly increased in AVMC patients compared to healthy volunteers(P<0.05).Moreover,the number of Th17 cells in peripheral blood CD4+T cells of AVMC patients showed a positive correlation with CVB3 virus titer(P<0.05).Following CVB3 infection,the number of Th17 cells increased compared with the control group(P<0.05),accompanied by elevated levels of IL-17,IL-21,and TNF-α in the supernatant(P<0.05).Knockdown of TLR7 and CVB3 infection in CD4+T cells significantly reduced the levels of Th17 cells(P<0.05),while the expression level of phosphorylated-activated TLR7 increased significantly after CVB3 infection of CD4+T cells compared to the control group(P<0.05).Treatment with the TLR7 inhibitor M5049 and CVB3 infection led to a significant decrease in Th17 cell levels(P<0.05).The secretion of IL-6 in CD4+T cells increased after CVB3 infection(P<0.05),and this increase was mitigated by TLR7 knockdown and CVB3 infection(P<0.05)as well as TLR7 inhibitor M5049 treatment and CVB3 infection(P<0.05).Conclusion:CVB3 activates TLR7 via phosphoryla-tion,prompting CD4+T cells to release IL-6 and undergo differentiation into Th17 cells.Consequently,TLR7 emerges as a promising therapeutic target for AVMC.

OBJECTIVE: To evaluate the functional and aesthetic evaluation of external fixator lengthening through plantar approach for fourth brachymetatarsia. METHODS: A retrospective analysis was conducted on 20 patients (23 feet) with fourth brachymetatarsia who met the selection criteria between January 2016 and January 2024, including 3 males and 17 females, with 8 left, 9 right, and 3 bilateral cases. The mean age was 24.7 years (range, 14-51 years). The preoperative metatarsal shortening length was (13.8±3.2) mm. The preoperative American Orthopaedic Foot and Ankle Society (AOFAS) forefoot score was 79.5±3.9, the visual analogue scale (VAS) score of appearance satisfaction was 1.7±0.8, and the appearance index (AI) score was 13.6±0.9. All patients underwent external fixator lengthening through plantar approach. The lengthening length of metatarsal bone, lengthening ratio, healing time, and healing index were recorded. Functional outcomes were assessed using the AOFAS forefoot score, VAS score of appearance satisfaction, and quality-of-life impact with AI questionnaire. RESULTS: All 20 patients were followed up 14-55 months with an average of 36.3 months. During the follow-up, complications occurred in 4 cases (17.4%), including 2 cases of metatarsophalangeal joint stiffness, which had no significant effect on the function and appearance. Delayed union of osteotomy occurred in 1 case (healed at 12 weeks after operation). Pin loosening occurred in 1 case and recovered after outpatient reinforcement. No complications related to plantar scar occurred. At last follow-up, the lengthening length of metatarsal bone was (13.9±3.1) mm, and the lengthening ratio was 25.8%±5.6%. All cases achieved bony union, with a mean healing time of (64.3±12.5) days and a healing index of (46.9±4.8) d/cm. At last follow-up, AOFAS score was 98.9±2.1, the VAS score of appearance satisfaction was 9.3±0.7, and the AI score was 0.6±0.8, which significantly improved when compared with those before operation ( t=27.398, P<0.001; t=32.994, P<0.001; t=56.135, P<0.001). CONCLUSION External fixator lengthening through plantar approach is a safe and effective technique for fourth brachymetatarsia, achieving satisfactory functional and aesthetic outcomes.
Humans ; Male ; Female ; Adult ; External Fixators ; Retrospective Studies ; Bone Lengthening/instrumentation* ; Middle Aged ; Metatarsal Bones/abnormalities* ; Adolescent ; Young Adult ; Treatment Outcome ; Patient Satisfaction ; Esthetics ; Osteotomy/methods* ; Foot Deformities, Congenital/surgery*

Traditional development of small protein scaffolds has relied on display technologies and mutation-based engineering, which limit sequence and functional diversity, thereby constraining their therapeutic and application potential. Protein design tools have significantly advanced the creation of novel protein sequences, structures, and functions. However, further improvements in design strategies are still needed to more efficiently optimize the functional performance of protein-based drugs and enhance their druggability. Here, we extended an evolution-based design protocol to create a novel minibinder, BindHer, against the human epidermal growth factor receptor 2 (HER2). It not only exhibits super stability and binding selectivity but also demonstrates remarkable properties in tissue specificity. Radiolabeling experiments with ^99mTc, ⁶⁸Ga, and ¹⁸F revealed that BindHer efficiently targets tumors in HER2-positive breast cancer mouse models, with minimal nonspecific liver absorption, outperforming scaffolds designed through traditional engineering. These findings highlight a new rational approach to automated protein design, offering significant potential for large-scale applications in therapeutic mini-protein development.

Objective To investigate the effects of TEK receptor tyrosine kinase(Tie2)L914F mutation on the biological behavior of vascular endothelial cells and the changes of related signaling pathways in patients with venous malformations.Methods Gene sequen-cing was used to detect Tie2-L914F mutations in venous malformations.HE staining and immunohistochemical staining were used to de-tect the expression of platelet derived growth factor subunit B gene(PDGFB)and α-smooth muscle actin(α-SMA)in venous malfor-mations caused by the mutation.Lentivirus overexpressing Tie2-wild type(Tie2-WT),Tie2-L914F and Tie2-GFP(green fluorescent protein,GFP)infected human umbilical vein endothelial cells(HUVECs).Real-time fluorescence quantitative PCR was used to detect the expression level of Tie2 in endothelial cells expressing exogenous Tie2 and Flag-tagged protein Tie2 protein was detected by western blotting to verify transfection efficiency.The proliferation,apoptosis,migration and tube-forming ability of the cells were determined by CCK-8,flow cytometry,Transwell migration assay and Matrigel matrix gel tube-forming assay.Western blotting was used to detect the expression levels of Protein Kinase B(PKB/AKT),FOXO1 and their phosphorylation,and the expression level of PDGFB was detec-ted by ELISA.Results The number of patients with venous malformations with L914F mutations was about 33.3％.HE and immuno-histochemical staining showed that the expressions of PDGFB and α-SMA were significantly down-regulated in venous malformation tis-sues with Tie2-L914F mutation,and were positively correlated with the decrease in cell coverage of the tube wall.Compared with Tie2-WT endothelial cells,the apoptosis number of Tie2-L914F endothelial cells was significantly reduced,while the proliferation and mi-gration ability was significantly increased,and the tube-forming ability was significantly decreased.Western blotting and ELISA showed that the phosphorylation levels of AKT and FOXO1 downstream of Tie2 signaling pathway in endothelial cells expressing Tie2-L914F were significantly increased,and the expression level of PDGFB was significantly decreased.Conclusion In venous malformations,Tie2-L914F mutation may downregulate the expression of PDGFB through AKT signaling pathway,which affects the biological behavior of vascular endothelial cells.

Objective To explore the role of protective function of Sestrin2(Sesn2)to mitochondria in alleviating cognitive dysfunction in mice with sepsis-associated encephalopathy(SAE).Methods 6-week-old male C57BL/6J mice were randomly divided into three groups:sham group,CLP group and CLP plus eupatilin group,40 mice in each group.A sepsis model was induced by cecal ligation and perforation(CLP).The CLP plus eupatilin group was treated with eupatilin.Neurobehavioral test and Morris water maze(MWM)were used to deter-mine neurobehavior and spatial learning and memory function in mice.The number of neurons in hippocampal CA1 area was counted by Nissl staining.HT22 cells were randomly divided into a control group(Con),lipopolysaccha-ride group(LPS),LPS plus eupatilin treatment group(LPS plus eupatilin)and LPS plus eupatilin and Nrf2 siRNA treatment group(LPS plus eupatilin and si-Nrf2).Apoptosis was analyzed by terminal deoxynucleotidyl transferase-mediated nick end labeling(TUNEL)staining,Mitochondrial membrane potential(MMP)was used to analyze mitochondrial damage.Results Seven days after CLP,as compared with sham mice,Sesn2 in hippocampus and cortex decreased significantly in CLP mice(P<0.01).As compared with CLP group,the survival rate in CLP plus eupatilin group increased significantly(P<0.05).As compared with sham group,the mice in CLP group showed a relatively high nerve injury score(P<0.05),and had fewer platform crossings and shorter target stay time,while the mice in CLP plus eupatilin group exhibited a lower injury score(P<0.05),and stayed in the target area for a longer time(P<0.05).As compared with sham group,the co-localization rate of neurons,Sesn2 and Nrf2 in CLP group decreased significantly(P<0.05),and the number of CD68/Iba-1 positive microglia increased significantly(P<0.05),while CLP plus eupatilin group reversed these changes.As compared with Con group,apoptosis and MMP level in LPS group increased significantly(P<0.01),while apoptosis and MMP level in LPS plus eupatilin group were lower than those in LPS group(P<0.05).However,Nrf2 knockdown(LPS plus eupatilin and si-Nrf2 group)reversed the anti-apoptosis and mitochondrial protection of eupatilin.Conclusions Eupatilin can alleviate cognitive dysfunction and neurological deficit in SAE mice by activating Sesn2-Nrf2 pathway,and improve inflammatory microenvironment by alleviating mitochondrial dysfunction.

The related symptoms of patients with congenital microtia after ear reconstruction surgery mainly include pain, psychological problems, fatigue, and sleep disorders. Measures of chest strap compression bandage, diversion of attention and use of analgesics; psychological counseling, establishment of WeChat communication group, and organizing communication meetings; proper lying position, earmuffs, emotional reassurance were taken to improve symptoms. This article provides a summary review of the management measures for related symptoms, aiming to draw more attention from healthcare professionals and to provide a reference for constructing symptom management programs in the future, promoting the patients’ physical and mental health recovery and enhancing their quality of life.

The related symptoms of patients with congenital microtia after ear reconstruction surgery mainly include pain, psychological problems, fatigue, and sleep disorders. Measures of chest strap compression bandage, diversion of attention and use of analgesics; psychological counseling, establishment of WeChat communication group, and organizing communication meetings; proper lying position, earmuffs, emotional reassurance were taken to improve symptoms. This article provides a summary review of the management measures for related symptoms, aiming to draw more attention from healthcare professionals and to provide a reference for constructing symptom management programs in the future, promoting the patients’ physical and mental health recovery and enhancing their quality of life.