Objective To investigate the mechanism by which Shenqi Xiaozheng Decoction-medicated serum regulates c-Myc/SLC1A5/GLS1 signaling axis to inhibit glutamine(Gln)metabolism and activate immunogenic cell death(ICD)in non-small cell lung cancer(NSCLC)cells.Methods A549 cells were divided into control group,model group,Shenqi Xiaozheng Decoction-medicated serum low-,medium-and high-dosage groups and positive control group.A Gln-dependent growth model was established,and cells were treated with different concentrations of Shenqi Xiaozheng Decoction-medicated serum or the SLC1A5 inhibitor V-9302.Cell proliferation was assessed by CCK-8,EdU and colony formation assays;Cell invasion and migration were evaluated using Transwell and wound-healing assays;intracellular Gln,glutathione(GSH),and α-ketoglutarate(α-KG)contents were determined by colorimetric assay;reactive oxygen species(ROS)contents were measured with fluorescent probes;Western blot was used to detect the protein expressions of E-cadherin,N-cadherin,c-Myc,SLC1A5 and GLS1;c-Myc/SLC1A5 colocalization and high mobility group box 1(HMGB1)expression were assessed by dual immunofluorescence staining;flow cytometry was used to evaluate calreticulin(CRT)exposure on the cell surface,and ATP and HMGB1 contents in cell supernatants were quantified by ELISA.Results Compared with the control group,the model group showed significantly increased A549 cell viability,EdU-positive rate and migration rate(P<0.05),as well as higher colony counts and invasion cell numbers(P<0.05);cellular Gln,GSH and α-KG contents were significantly elevated(P<0.05,P<0.01),while ROS content were not significantly different(P>0.05),E-cadherin protein expression significantly decreased,whereas the protein expressions of N-cadherin,c-Myc,SLC1A5 and GLS1 significantly increased(P<0.05,P<0.01).c-Myc and SLC1A5 colocalization was enhanced,HMGB1 expression was significantly increased(P<0.01),CRT exposure significantly increased(P<0.01),and ATP and HMGB1 contents in cell supernatant were significantly elevated(P<0.05,P<0.01).Compared with the model group,Shenqi Xiaozheng Decoction-medicated serum at different concentrations significantly inhibited Gln-stimulated A549 cell proliferation,migration and invasion in a dosage-dependent manner.Mechanistic studies indicated that Shenqi Xiaozheng Decoction could reduce Gln uptake and synthesis of its metabolic products GSH and α-KG,induce ROS accumulation,up-regulate protein expression of E-cadherin,down-regulate the protein expressions of N-cadherin,c-Myc,SLC1A5 and GLS1(P<0.05,P<0.01),and enhance CRT exposure,ATP release and HMGB1 secretion(P<0.01).Conclusion Shenqi Xiaozheng Decoction may exert a synergistic"metabolism-immunity"antitumor effect by inhibiting c-Myc/SLC1A5/GLS1 axis-mediated Gln uptake,inducing ROS accumulation,and activating ICD signaling.

Objective To evaluate the efficacy and safety of Qixian Tongluo Formula in the treatment of post-cerebral infarction paralysis with kidney deficiency and blood stasis syndrome,and to preliminarily explore the molecular mechanism of Qixian Tongluo Formula in improving impaired motor function from the perspective of cross-kingdom regulation of Chinese medicine microRNA(miRNA).Methods A pragmatic randomized controlled trial was conducted with 102 patients in the recovery period of post-cerebral infarction paralysis with kidney deficiency and blood stasis syndrome in our hospital.Patients were randomly divided into trial group and control group,with 51 cases in each group.The control group received standard Western medicine standard treatment,while the trial group received Qixian Tongluo Formula in addition to the standard treatment,with one dose per day,boiled in water,and taken warm after breakfast and dinner for a course of 2 months.The disability rate was used as the main efficacy indicator,and the incidence of adverse reactions was used as a safety indicator.miRNA from patient serum and Qixian Tongluo decoction were extracted respectively,and high-throughput sequencing was performed.The two sequences were compared to screen out the cross-kingdom gene transfer of Chinese medicine miRNA.Finally,its target genes of miRNA were predicted,and GO function and KEGG pathway enrichment analysis were carried out.Results A total of 67 patients completed the clinical trial,including 36 cases in the trial group and 31 cases in the control group;The disability rate in the trial group(13.9%)was lower than that in the control group(35.5%)(P<0.05);The incidence of adverse reactions was similar between the trial group(7.69%)and the control group(6.06%)(P>0.05);A total of 9530 Qixian Tongluo decoction miRNA sequences were screened,with 150 potentially involved in cross-kingdom gene transfer,including families such as miR-15 and miR-17;According to the target gene prediction of the top 10 miRNAs in cross-kingdom gene transfer of Chinese medicine,345 overlapping target genes were obtained;GO functional enrichment analysis revealed 16 biological processes,7 cellular components,and 2 molecular functions among the top 25 enriched functions,while KEGG pathway analysis mainly focused on the transforming growth factor-βsignaling pathway,neurotrophin signaling pathway,which are closely related to neural repair and functional recovery processes such as glial scar formation and synaptic plasticity after cerebral ischemia.Conclusion Qixian Tongluo Formula can significantly improve the functional independence level of patients with kidney deficiency and blood stasis syndrome in the recovery period of paralysis after cerebral infarction,offering a safe and effective treatment option for these patients;There were a large number of miRNAs in Qixian Tongluo decoction,some of which could cross-kingdom transferred into the human blood circulation,and promote the recovery of motor function in patients with cerebral infarction through multi-target,multi link and multi pathway gene network regulation.This study provides a new idea for subsequent clinical and basic research.

Objective To evaluate the efficacy and safety of Qixian Tongluo Formula in the treatment of post-cerebral infarction paralysis with kidney deficiency and blood stasis syndrome,and to preliminarily explore the molecular mechanism of Qixian Tongluo Formula in improving impaired motor function from the perspective of cross-kingdom regulation of Chinese medicine microRNA(miRNA).Methods A pragmatic randomized controlled trial was conducted with 102 patients in the recovery period of post-cerebral infarction paralysis with kidney deficiency and blood stasis syndrome in our hospital.Patients were randomly divided into trial group and control group,with 51 cases in each group.The control group received standard Western medicine standard treatment,while the trial group received Qixian Tongluo Formula in addition to the standard treatment,with one dose per day,boiled in water,and taken warm after breakfast and dinner for a course of 2 months.The disability rate was used as the main efficacy indicator,and the incidence of adverse reactions was used as a safety indicator.miRNA from patient serum and Qixian Tongluo decoction were extracted respectively,and high-throughput sequencing was performed.The two sequences were compared to screen out the cross-kingdom gene transfer of Chinese medicine miRNA.Finally,its target genes of miRNA were predicted,and GO function and KEGG pathway enrichment analysis were carried out.Results A total of 67 patients completed the clinical trial,including 36 cases in the trial group and 31 cases in the control group;The disability rate in the trial group(13.9%)was lower than that in the control group(35.5%)(P<0.05);The incidence of adverse reactions was similar between the trial group(7.69%)and the control group(6.06%)(P>0.05);A total of 9530 Qixian Tongluo decoction miRNA sequences were screened,with 150 potentially involved in cross-kingdom gene transfer,including families such as miR-15 and miR-17;According to the target gene prediction of the top 10 miRNAs in cross-kingdom gene transfer of Chinese medicine,345 overlapping target genes were obtained;GO functional enrichment analysis revealed 16 biological processes,7 cellular components,and 2 molecular functions among the top 25 enriched functions,while KEGG pathway analysis mainly focused on the transforming growth factor-βsignaling pathway,neurotrophin signaling pathway,which are closely related to neural repair and functional recovery processes such as glial scar formation and synaptic plasticity after cerebral ischemia.Conclusion Qixian Tongluo Formula can significantly improve the functional independence level of patients with kidney deficiency and blood stasis syndrome in the recovery period of paralysis after cerebral infarction,offering a safe and effective treatment option for these patients;There were a large number of miRNAs in Qixian Tongluo decoction,some of which could cross-kingdom transferred into the human blood circulation,and promote the recovery of motor function in patients with cerebral infarction through multi-target,multi link and multi pathway gene network regulation.This study provides a new idea for subsequent clinical and basic research.

Objective:To compare the biomechanical differences among modular total sacral prosthesis, integrated total sacral prosthesis and screw-rod system for lumbosacral reconstruction after total sacrectomy by finite element analysis.Methods:Three finite element models of reconstruction after total sacrectomy were established: six-rod plus anterior column, integrated total sacral prosthesis, and modular total sacral prosthesis. A vertical load of 600 N was applied to the L 3 vertebra, and the bilateral acetabula were fixed in all degrees of freedom to restrict their movement, simulating a bipedal standing posture. The maximum stress, stress distribution on the iliac screws, stress distribution on the longitudinal rods, the shift-down displacement of the L 5 vertebra, and the stress direction on the contact surface between the prosthesis and the ilium on all implant components (including prosthesis, screws, and connecting rods) were compared. Results:Finite element analysis results show that the average maximum stress of the six-rod plus anterior column reconstruction on all implant instrumentation was 217.9±10.2 MPa, the integrated total sacral prosthesis reconstruction was 185.7±21.1 MPa, and the modular total sacral prosthesis reconstruction was 157.4±31.2 MPa. The differences were statistically significant ( F=12.357, P<0.001). Among them, the difference between the modular total sacral prosthesis reconstruction and the six-rod plus anterior column reconstruction was statistically significant ( P<0.001), while the difference between the modular total sacral prosthesis reconstruction and the integrated total sacral prosthesis reconstruction was not statistically significant ( P=0.051). The maximum stress on the iliac bone screws and longitudinal connecting rods: for the six-rod plus anterior column reconstruction, it was 157.2 MPa and 105.4 MPa respectively; for the integrated total sacral prosthesis reconstruction, it was 59.2 MPa and 97.8 MPa respectively; for the modular total sacral prosthesis reconstruction, it was 58.4 MPa and 35.6 MPa respectively. The distance of L 5 vertebral body downward displacement: for the six-rod plus anterior column reconstruction, it was 1.05±0.06 mm; for the integrated total sacral prosthesis reconstruction, it was 0.34±0.02 mm; for the modular total sacral prosthesis reconstruction, it was 0.40±0.05 mm. The difference was statistically significant ( F=357.730, P<0.001), among which the differences between the modular total sacral prosthesis reconstruction and the six-rod plus anterior column reconstruction and that between the integrated total sacral prosthesis reconstruction and the six-rod plus anterior column reconstruction were all statistically significant ( P<0.05), while the difference between the modular total sacral prosthesis reconstruction and the integrated total sacral prosthesis reconstruction was not statistically significant ( P=0.145). The stress on the iliac bone contact surface of the integrated total sacral prosthesis was 34.2° and manifested as shear force; the stress on the iliac bone contact surface of the modular total sacral prosthesis was 88.9° and manifested as compressive stress. Conclusions:This modular total sacral prosthesis exhibits lower peak stress compared with the integrated total sacral prosthesis and screw-rod system. The spinal stability of the modular total sacral prosthesis is comparable to that of the integrated total sacral prosthesis and superior to that of the screw-rod system.

Objective To investigate the effect of extracellular heat shock protein(HSP)22 on Toll-like receptor(TLR)4/nuclear factor-κB(NF-κB)signaling pathway in oxidative-low-density lipoprotein(ox-LDL)-induced inflammatory damage in coronary endothelial cells(HCAECs).Methods HCAECs were cul-tured in vitro and pretreated with ox-LDL to establish a model of high-lipid-induced endothelial cell injury.Re-combinant human HSP22(rhHSP22)was exogenously treated.The effects of rhHSP22 on the expression of inflammation-related proteins such as interleukin(IL)-8,vascular cell adhesion molecule(VCAM)-1 and NF-κB in endothelial cells and endothelial cell apoptosis were observed.The relationship between HSP22 and TLR4/NF-κB signaling pathway was investigated under the action of TLR4 inhibitor E5564.Western blot was used to detect the expression of IL-8,VACM-1 and NF-κB proteins,and flow cytometry was used to detect the apoptosis of endothelial cells in each group.Results Compared with the CNT group,the relative expression levels of IL-8,VACM-1 and NF-κB protein in the rhHSP22 group,rhHSP22+ox-LDL group,rhHSP22+E5564 group and rhHSP22+E5564+ox-LDL group were significantly increased,and the differences were sta-tistically significant(P<0.05).Compared with the rhHSP22 group,the relative expression levels of IL-8,VACM-1 and NF-κB protein in the rhHSP22+ox-LDL group were significantly increased,and the differences were statistically significant(P<0.05).Compared with the rhHSP22+ox-LDL group,the relative expression levels of IL-8 and VACM-1 in the rhHSP22+E5564+ox-LDL group were decreased,and the differences were statistically significant(P<0.05).Compared with the CNT group,the apoptosis rate in the rhHSP22 group,rhHSP22+ox-LDL group and rhHSP22+E5564+ox-LDL group was significantly increased,and the differ-ence was statistically significant(P<0.05).Compared with the rhHSP22 group,the apoptosis rate in the rhHSP22+ox-LDL group was increased,and the difference was statistically significant(P<0.05).Compared with the rhHSP22+ox-LDL group,the apoptosis rate in the rhHSP22+E5564+ox-LDL group was de-creased,and the difference was statistically significant(P<0.05).Conclusion In ox-LDL-induced inflamma-tory damage of HCAECs,extracellular HSP22 induces the expression of IL-8,VACM-1 and NF-κB proteins by activating the TLR4/NF-κB signaling pathway,and promotes endothelial cell apoptosis.

Objective:To compare the biomechanical differences among modular total sacral prosthesis, integrated total sacral prosthesis and screw-rod system for lumbosacral reconstruction after total sacrectomy by finite element analysis.Methods:Three finite element models of reconstruction after total sacrectomy were established: six-rod plus anterior column, integrated total sacral prosthesis, and modular total sacral prosthesis. A vertical load of 600 N was applied to the L 3 vertebra, and the bilateral acetabula were fixed in all degrees of freedom to restrict their movement, simulating a bipedal standing posture. The maximum stress, stress distribution on the iliac screws, stress distribution on the longitudinal rods, the shift-down displacement of the L 5 vertebra, and the stress direction on the contact surface between the prosthesis and the ilium on all implant components (including prosthesis, screws, and connecting rods) were compared. Results:Finite element analysis results show that the average maximum stress of the six-rod plus anterior column reconstruction on all implant instrumentation was 217.9±10.2 MPa, the integrated total sacral prosthesis reconstruction was 185.7±21.1 MPa, and the modular total sacral prosthesis reconstruction was 157.4±31.2 MPa. The differences were statistically significant ( F=12.357, P<0.001). Among them, the difference between the modular total sacral prosthesis reconstruction and the six-rod plus anterior column reconstruction was statistically significant ( P<0.001), while the difference between the modular total sacral prosthesis reconstruction and the integrated total sacral prosthesis reconstruction was not statistically significant ( P=0.051). The maximum stress on the iliac bone screws and longitudinal connecting rods: for the six-rod plus anterior column reconstruction, it was 157.2 MPa and 105.4 MPa respectively; for the integrated total sacral prosthesis reconstruction, it was 59.2 MPa and 97.8 MPa respectively; for the modular total sacral prosthesis reconstruction, it was 58.4 MPa and 35.6 MPa respectively. The distance of L 5 vertebral body downward displacement: for the six-rod plus anterior column reconstruction, it was 1.05±0.06 mm; for the integrated total sacral prosthesis reconstruction, it was 0.34±0.02 mm; for the modular total sacral prosthesis reconstruction, it was 0.40±0.05 mm. The difference was statistically significant ( F=357.730, P<0.001), among which the differences between the modular total sacral prosthesis reconstruction and the six-rod plus anterior column reconstruction and that between the integrated total sacral prosthesis reconstruction and the six-rod plus anterior column reconstruction were all statistically significant ( P<0.05), while the difference between the modular total sacral prosthesis reconstruction and the integrated total sacral prosthesis reconstruction was not statistically significant ( P=0.145). The stress on the iliac bone contact surface of the integrated total sacral prosthesis was 34.2° and manifested as shear force; the stress on the iliac bone contact surface of the modular total sacral prosthesis was 88.9° and manifested as compressive stress. Conclusions:This modular total sacral prosthesis exhibits lower peak stress compared with the integrated total sacral prosthesis and screw-rod system. The spinal stability of the modular total sacral prosthesis is comparable to that of the integrated total sacral prosthesis and superior to that of the screw-rod system.

Objective To investigate the mechanism by which Shenqi Xiaozheng Decoction-medicated serum regulates c-Myc/SLC1A5/GLS1 signaling axis to inhibit glutamine(Gln)metabolism and activate immunogenic cell death(ICD)in non-small cell lung cancer(NSCLC)cells.Methods A549 cells were divided into control group,model group,Shenqi Xiaozheng Decoction-medicated serum low-,medium-and high-dosage groups and positive control group.A Gln-dependent growth model was established,and cells were treated with different concentrations of Shenqi Xiaozheng Decoction-medicated serum or the SLC1A5 inhibitor V-9302.Cell proliferation was assessed by CCK-8,EdU and colony formation assays;Cell invasion and migration were evaluated using Transwell and wound-healing assays;intracellular Gln,glutathione(GSH),and α-ketoglutarate(α-KG)contents were determined by colorimetric assay;reactive oxygen species(ROS)contents were measured with fluorescent probes;Western blot was used to detect the protein expressions of E-cadherin,N-cadherin,c-Myc,SLC1A5 and GLS1;c-Myc/SLC1A5 colocalization and high mobility group box 1(HMGB1)expression were assessed by dual immunofluorescence staining;flow cytometry was used to evaluate calreticulin(CRT)exposure on the cell surface,and ATP and HMGB1 contents in cell supernatants were quantified by ELISA.Results Compared with the control group,the model group showed significantly increased A549 cell viability,EdU-positive rate and migration rate(P<0.05),as well as higher colony counts and invasion cell numbers(P<0.05);cellular Gln,GSH and α-KG contents were significantly elevated(P<0.05,P<0.01),while ROS content were not significantly different(P>0.05),E-cadherin protein expression significantly decreased,whereas the protein expressions of N-cadherin,c-Myc,SLC1A5 and GLS1 significantly increased(P<0.05,P<0.01).c-Myc and SLC1A5 colocalization was enhanced,HMGB1 expression was significantly increased(P<0.01),CRT exposure significantly increased(P<0.01),and ATP and HMGB1 contents in cell supernatant were significantly elevated(P<0.05,P<0.01).Compared with the model group,Shenqi Xiaozheng Decoction-medicated serum at different concentrations significantly inhibited Gln-stimulated A549 cell proliferation,migration and invasion in a dosage-dependent manner.Mechanistic studies indicated that Shenqi Xiaozheng Decoction could reduce Gln uptake and synthesis of its metabolic products GSH and α-KG,induce ROS accumulation,up-regulate protein expression of E-cadherin,down-regulate the protein expressions of N-cadherin,c-Myc,SLC1A5 and GLS1(P<0.05,P<0.01),and enhance CRT exposure,ATP release and HMGB1 secretion(P<0.01).Conclusion Shenqi Xiaozheng Decoction may exert a synergistic"metabolism-immunity"antitumor effect by inhibiting c-Myc/SLC1A5/GLS1 axis-mediated Gln uptake,inducing ROS accumulation,and activating ICD signaling.

The timely use of screening tools to assess the palliative care needs of emergency patients can provide decision-making basis for patient referral and help patients receive palliative care in a timely manner.This paper reviews the palliative care needs screening tools for emergency patients at home and abroad,describes their development methods,scoring criteria and application status,compares and analyzes their characteristics and shortcomings.It aims to provide references for the localized development and application of palliative care needs screening tools in emergency patients.

Objective To investigate the role of sodium-glucose cotransporter 1(SGLT1)inhibitor mizagliflozin(MIZA)in autosomal dominant polycystic kidney disease(ADPKD).Methods Western blotting,quantitative polymerase chain reaction(qPCR),and immunofluorescence staining were used to determine the expression and distribution of SGLT1 in kidney tissues of PKD1-/-and PKD1+/+mice,human renal cancer adjacent tissue and ADPKD tissue.Renal cyst lining epithelial cells OX161 and renal tubular epithelial cells UCL93 were treated with MIZA,incubated at 37℃for 24,48,and 72 h,and then were subjected to methyl thiazolyl tetrazolium and colony formation assay to observe cell proliferation.The qPCR method was used to determine the mRNA levels of collagen 1α1,collagen 3α1,and fibronectin 1 in OX161 cells treated with 100 μmol/L MIZA for 48 h.The Madin-Darby canine kidney(MDCK)cell 3D cyst formation assay verified the effect of MIZA on cyst formation.The mRNA-seq technology was used to detect differentially expressed genes between UCL93 cells and OX161 cells,and between OX161 cells and OX161 cells treated with 100 μmol/L MIZA for 48 h,and then the differentially expressed genes were analyzed with Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.Results The expression level of SGLT1 was significantly increased in the tissues of ADPKD patients and PKD1-/-mice compared to those in normal kidney tissues(P＜0.05,P＜0.01).Immunofluorescence staining revealed that SGLT1 was mainly expressed in the cystic lining epithelial cells.Additionally,MIZA inhibited the proliferation and fibrosis of polycystic kidney cells in a concentration-and time-dependent manner,and also inhibited cyst formation in 3D formation assay in vitro.The mRNA-seq analysis and KEGG enrichment analysis showed that differentially expressed genes between OX161 cells and OX161 cells cultured in 100 μmol/L MIZA for 48 h were mainly enriched in the phosphatidylinositol 3-kinase(PI3K)-protein kinase B(Akt)and mitogen-activated protein kinase(MAPK)signaling pathways,which were the same as those between OX161 cells and UCL93 cells.Conclusion The SGLT1 inhibitor MIZA may inhibit the proliferation and fibrosis of polycystic kidney cells through signaling pathways such as PI3K-Akt and MAPK,delaying the growth of polycystic kidney,and it is a potential therapeutic target for ADPKD.

BACKGROUND:Numerous scholars have previously researched certain greater tuberosity fractures and the procedures used to treat them.Few researchers,however,have studied the comminuted split fracture of the greater tuberosity of the humerus(Liu-Gang type IV)with rotator cuff tear in great detail. OBJECTIVE:To compare the clinical therapeutic effect of open repair position modified calcaneal plate combined with suture anchors and proximal humeral internal locking system(PHILOS)plate in the treatment of comminuted fracture of split-type greater tuberosity of humerus combined with rotator cuff tears(Liu-Gang type IV). METHODS:Case data of 30 patients with comminuted fracture of split-type greater tuberosity of humerus combined with rotator cuff tears(Liu-Gang type IV)from May 2012 to May 2022 were retrospectively analyzed.They were divided into the modified calcaneal plate combined with suture anchor group(group A)and the PHILOS with#2 Johnson group(group B),with 15 cases in each group.Intraoperative blood loss,surgical time,and incision length of all patients were recorded.Pain visual analog scale score,Constant-Murley score,as well as shoulder joint abduction,forward flexion,external rotation,and dorsal expansion activities during the last follow-up(>1 year)were evaluated. RESULTS AND CONCLUSION:(1)The surgical incision length and operation time were shorter,and blood loss was less in group A than those in group B(P<0.05).(2)No significant difference in visual analog scale score and Constant-Murley score was detected between the two groups(P>0.05).(3)During the last follow-up,forward flexion in group A was better than that in group B(P<0.05).No significant difference in abduction,external rotation,and dorsal expansion was determined between group A and group B(P>0.05).(4)In terms of complications,there was 1 case of shoulder joint pain and discomfort in group A(7%),2 cases of subacromial impingement syndrome,2 cases of upward movement of nodules,and 2 cases of shoulder joint pain(40%)in group B.There were significant differences in complication rates between the two groups(P=0.031).(5)In summary,the modified calcaneal plate combined with suture anchors in the treatment of comminuted fracture of split-type greater tuberosity of humerus combined with rotator cuff tears(Liu-Gang type IV)could better restore the forward flexion function of the shoulder joint and has a small incision,less blood loss,shorter operation time and fewer complications.