BACKGROUND:The elastic modulus of bone-cartilage integration scaffolds differs significantly from that of natural bone-cartilage tissue,which can lead to a stress shield effect.As a result,the implants become loose and deformed,affecting the repair of osteochondral tissue.Cell gradient scaffolds made by axial direction three-period minimal surface have the same porosity and elasticity modulus as the human body,which provides a new idea for bone-cartilage scaffold design.OBJECTIVE:To study the effect of cell type and pore size on the mechanical properties of cell gradient scaffolds.METHODS:Three basic cells of Gyroid(G)type,Diamond(D)type,and Primitive(P)type were used.Through mathematical modeling of three-period minimal surface,different sizes and types of cells were used in the gradient region.A total of six kinds of cell gradient scaffolds(G-2P-4D,P-2D-4G,D-2P-4D,G-2D-4P,P-2G-4D,and D-2G-4P)were constructed and mechanical experiments and simulation experiments were conducted to evaluate the mechanical properties of the scaffolds.Flow performance parameters of the fluids in the scaffolds were obtained through computational fluid dynamics simulation.RESULTS AND CONCLUSION:Finite element mechanical simulation and compression experiment showed that P-2G-4D and P-2D-4G with the highest elastic modulus(148.67 MPa and 152.1 MPa),bearing a higher body load,improved the stability of the scaffold.The stress distribution in D-2P-4G was even and effectively reduced stress concentration,so that the connection function area could effectively transfer stress and reduce stress shielding.Flow rate was changing the least in G-2D-4P(0.10-0.48 mm/s).Permeability was higher than other scaffolds so that body fluids were able to flow though the gradient scaffold after implantation.This design method provides a new idea for the design of osteochondral scaffolds,and the simulation analysis results also provide a reference for the prediction of bone integration after implantation of scaffolds.

Objective:To construct a model of mitochondria-related genes (Mito-RGs) in hepatocellular carcinoma (HCC), and predict the immune, prognostic and therapeutic characteristics of HCC based on the model, so as to provide a new idea for the diagnosis and treatment of HCC.Methods:The expression profiles of HCC and corresponding clinical information were obtained from the Cancer Genome Atlas (TCGA) database. Univariate Cox regression, Least Absolute Shrinkage and Selection Operator (LASSO) regression and multivariate Cox regression were used to construct a prognostic model of HCC based on Mito-RGs, and the International Cancer Genome Consortium-Liver Cancer-RIKEN-Japan ICGC-LIRI-JP dataset were used for validation. GO and KEGG analyses were performed to investigate the signaling pathways enriched for differentially expressed genes in the high- and low-risk groups. Immune infiltration was assessed using CIBERSORT. Single-cell data were used to study the proportion of immune cells in high- and low-risk groups of HCC samples and the relationship with cell proliferation. Cellminer was used to assess the relationship between risk score models and drug sensitivity.Results:A risk-prognostic model of HCC containing seven Mito-RGs ( DTYMK, ACADS, HMGCS2, CYP27A1, TOMM40L, STOM, and AKR1B10) was constructed. High-risk HCC patients had a worse prognosis. Genes upregulated in the high- and low-risk groups of differentially expressed genes were enriched in cell cycle and metabolism-related pathways. Single-cell data showed higher proportions of CD8 + T cells, macrophages and monocytes, and proliferating cells in the high-risk group. CIBERSORT analysis suggested that Treg cells and M0 macrophages were more abundant in the high-risk group, whereas CD8 + T cells and CD4 + memory T cells were less abundant. Patients in the high-risk group were more sensitive to myeloid cell leukemia sequence 1 inhibitor, vincristine, phosphatidylinositol kinase beta subunit inhibitor, and aurora kinase A, while trametinib, selumetinib, extracellular regulated protein kinase, and mitogen-activated extracellular signal-regulated kinase were more effective in the low-risk group. Conclusion:The constructed Mito-RGs model is capable of providing a more accurate assessment of the prognosis and the degree of immune cell infiltration in HCC patients.

ObjectiveTo evaluate the effects of Tongnaoyin on the blood-brain barrier status and neurological impairment in acute ischemic stroke （AIS） patients with the syndrome of phlegm-stasis blocking collaterals by dynamic contrast-enhanced magnetic resonance imaging （DCE-MRI）. MethodsA total of 63 patients diagnosed with AIS in the Jiangsu Province Hospital of Chinese Medicine from October 2022 to December 2023 were enrolled in this study. According to random number table method，the patients were assigned into a control group （32 cases） and an observation group （31 cases）. The control group received conventional Western medical treatment，and the observation group took 200 mL Tongnaoyin after meals，twice a day from day 2 of admission on the basis of the treatment in the control group. After 7 days of treatment，the patients were examined by DCE-MRI. The baseline data for two groups of patients before treatment were compared. The National Institute of Health Stroke Scale （NIHSS） score and modified Rankin Scale （mRS） score were recorded before treatment and after 90 days of treatment for both groups. The rKtrans，rKep，and rVe values were obtained from the region of interest （ROI） of the infarct zone/mirror area and compared between the two groups. ResultsThere was no significant difference in the NIHSS or mRS score between the two groups before treatment. After 90 days of treatment，the NIHSS and mRS scores declined in both groups，and the observation group had lower scores than the control group （P<0.05）. After treatment，the rKtrans and rVe in the observation group were lower than those in the control group （P<0.01）. ConclusionCompared with conventional Western medical treatment alone，conventional Western medical treatment combined with Tongnaoyin accelerates the repair of the blood-brain barrier in AIS patients，thereby ameliorating neurological impairment after AIS to improve the prognosis.

OBJECTIVE: To observe the clinical efficacy of Daoqi (directing qi flowing) acupuncture technique in Huangdi Neijing (Yellow Emperor's Inner Classic) for moderate-to-severe obstructive sleep apnea-hypopnea syndrome (OSAHS). METHODS: Sixty patients with moderate-to-severe OSAHS were randomly divided into a Daoqi acupuncture group (30 cases) and a conventional acupuncture group (30 cases, 1 case dropped out). In the Daoqi acupuncture group, Daoqi acupuncture technique in Huangdi Neijing was applied at Shanglianquan (Extra), Fengfu (GV16), and bilateral Lieque (LU7), Zhaohai (KI6); in the conventional acupuncture group, conventional acupuncture was applied at Shanglianquan (Extra), Fengfu (GV16), Yamen (GV15), and bilateral Lieque (LU7), Zhaohai (KI6), Zusanli (ST36), Fenglong (ST40). The treatment was adopted once every other day, 3 times a week, 4 weeks as one course and 3 courses were required totally in both groups. Before and after treatment, the polysomnography (PSG) indexes [apnea-hypopnea index (AHI), hypopnea index (HI), apnea index (AI), longest apnea duration, lowest nocturnal SaO₂ (LSaO₂)], and scores of Epworth sleepiness scale (ESS), Pittsburgh sleep quality index (PSQI), World Health Organization quality of life-BREF (WHOQOL-BREF) were observed, and the clinical efficacy was evaluated after treatment in the two groups. RESULTS: After treatment, the AHI, HI, AI and longest apnea duration were reduced compared with those before treatment in the two groups (P<0.01), the LSaO₂ was increased in the Daoqi acupuncture group (P<0.01); in the Daoqi acupuncture group, the AHI, HI, AI and longest apnea duration were lower than those in the conventional acupuncture group (P<0.05), and the LSaO₂ was higher than that in the conventional acupuncture group (P<0.05). After treatment, the ESS and PSQI scores were decreased compared with those before treatment (P<0.01), while the WHOQOL-BREF scores were increased compared with those before treatment (P<0.01) in the two groups; in the Daoqi acupuncture group, the ESS and PSQI scores were lower than those in the conventional acupuncture group (P<0.05, P<0.01), and the WHOQOL-BREF score was higher than that in the conventional acupuncture group (P<0.05). The total effective rate was 93.3% (28/30) in the Daoqi acupuncture group, which was higher than 82.8% (24/29) in the conventional acupuncture group (P<0.01). CONCLUSION Daoqi acupuncture technique in Huangdi Neijing can effectively treat moderate-to-severe OSAHS patients, improve the clinical symptoms and quality of life, and has the advantages i.e. simpler acupoints selection and gentler stimulation.
Humans ; Sleep Apnea, Obstructive/physiopathology* ; Male ; Female ; Middle Aged ; Acupuncture Therapy ; Adult ; Acupuncture Points ; Treatment Outcome ; Aged ; Quality of Life

In order to establish a serological method for the detection of bovine respiratory syncytial virus,the prokaryotic expression of four proteins of BRSV,G,F,P,and M was carried out,and the most suitable coating antigen was screened to establish an indirect ELISA detection method.The results showed that the four recombinant proteins of BRSV,rG,rF,rP and rM were successfully expressed.The results of checkerboard screening showed that the P/N value of rG protein was the largest,which was determined to be the best coating antigen established by indirect ELISA meth-od.The optimal reaction conditions for indirect ELISA were as follows:the mass concentration of rG protein coating was 1 mg/L,37℃ for 2 h;3％BSA 37℃ block for 1 h;Serum was diluted 1∶50 and incubated at 37℃ for 1h;Secondary antibody 1∶5 000 dilution,37℃ for 30min;The color development conditions of the substrate were 37℃ for 15 min;Thirty negative sera were selected,and the cut-off value was determined to be 0.63 by the established indirect ELISA method.The re-sults of the specificity test showed that the indirect ELISA method established in this test only recognized BRSV-positive serum,and did not react with IBRV,BCoV,and BPIV3-positive serum.The results of repeatability test showed that the method had good repeatability,and the coefficient of variation within and between batches was less than 10％.The results of the sensitivity test showed that the BRSV-positive serum was still positive when diluted to 1∶8 192.The indirect ELISA method established in this experiment was used to detect 100 clinical serum samples at the same time,and the total coincidence rate of the two reached 90.48％,the positive coincidence rate was 93.42％,and the negative coincidence rate was 82.75％.The indirect ELISA established in this test can be used for the detection of bovine respiratory syncytial virus in clinical practice.

Objective:To investigate the impact of the number of examined lymph nodes (ELN) on survival outcomes in gastric cancer patients with postoperative pathological stage pN3b.Methods:This retrospective cohort study included 279 pN3b gastric cancer patients who underwent D2 gastrectomy at Nanfang Hospital, Southern Medical University (September 2008 to April 2023), with 35 patients receiving combination chemotherapy and anti-PD-1 therapy (immunotherapy group) and 244 receiving adjuvant chemotherapy alone (nonimmunotherapy group). Additionally, 422 patients with pN3b from the SEER database (2005 to 2020) were collected as an external validation cohort to determine the optimal cutoff value for the number of lymph nodes examined in the nonimmunotherapy group. The primary endpoints were overall survival (OS) and recurrence-free survival (RFS) in the nonimmunotherapy group of the Nanfang Hospital cohort, stratified by whether the number of examined lymph nodes was above or below the ELN optimal cutoff value. These findings were subsequently validated in the SEER cohort.Results:The optimal ELN cutoff value (34 nodes) was determined using X-tile software and by constructing an ELN-HR fitting model with inflection point identification. In the nonimmunotherapy group, patients with ELN >34 exhibited significantly prolonged survival compared to ELN ≤34 (median OS: 25.0 (95%CI:20.5-29.5) to 17.0 (95%CI:12.7-21.3) months, P=0.004; median RFS: 19.0 (95%CI:15.6-22.4) to 13.0 (95%CI:9.5-16.5) months, P=0.048). Multivariate Cox analysis also showed ELN >34 to be an independent protective factor for both OS (HR=0.576, 95%CI: 0.397-0.836) and RFS (HR=0.701, 95%CI: 0.492-0.998). In the SEER cohort, ELN >34 was associated with a 5-month OS extension (19 to 14 months, P=0.065), with multivariate analysis supporting its independent prognostic significance (HR=0.729, 95%CI: 0.580-0.915, P=0.006). Notably, in the immunotherapy group, patients with ELN >34 ( n=30) achieved a median OS of 41 months, but the median OS had not been reached in the ELN ≤34 group ( n=5) (1 death at 48 months). Conclusion:Higher ELN (>34) correlates with improved survival in nonimmunotherapy-treated pN3b gastric cancer patients. However, in pN3b gastric cancer patients treated with immunotherapy, the optimal ELN threshold requires further exploration to determine.

In order to establish a serological method for the detection of bovine respiratory syncytial virus,the prokaryotic expression of four proteins of BRSV,G,F,P,and M was carried out,and the most suitable coating antigen was screened to establish an indirect ELISA detection method.The results showed that the four recombinant proteins of BRSV,rG,rF,rP and rM were successfully expressed.The results of checkerboard screening showed that the P/N value of rG protein was the largest,which was determined to be the best coating antigen established by indirect ELISA meth-od.The optimal reaction conditions for indirect ELISA were as follows:the mass concentration of rG protein coating was 1 mg/L,37℃ for 2 h;3％BSA 37℃ block for 1 h;Serum was diluted 1∶50 and incubated at 37℃ for 1h;Secondary antibody 1∶5 000 dilution,37℃ for 30min;The color development conditions of the substrate were 37℃ for 15 min;Thirty negative sera were selected,and the cut-off value was determined to be 0.63 by the established indirect ELISA method.The re-sults of the specificity test showed that the indirect ELISA method established in this test only recognized BRSV-positive serum,and did not react with IBRV,BCoV,and BPIV3-positive serum.The results of repeatability test showed that the method had good repeatability,and the coefficient of variation within and between batches was less than 10％.The results of the sensitivity test showed that the BRSV-positive serum was still positive when diluted to 1∶8 192.The indirect ELISA method established in this experiment was used to detect 100 clinical serum samples at the same time,and the total coincidence rate of the two reached 90.48％,the positive coincidence rate was 93.42％,and the negative coincidence rate was 82.75％.The indirect ELISA established in this test can be used for the detection of bovine respiratory syncytial virus in clinical practice.

BACKGROUND:The elastic modulus of bone-cartilage integration scaffolds differs significantly from that of natural bone-cartilage tissue,which can lead to a stress shield effect.As a result,the implants become loose and deformed,affecting the repair of osteochondral tissue.Cell gradient scaffolds made by axial direction three-period minimal surface have the same porosity and elasticity modulus as the human body,which provides a new idea for bone-cartilage scaffold design.OBJECTIVE:To study the effect of cell type and pore size on the mechanical properties of cell gradient scaffolds.METHODS:Three basic cells of Gyroid(G)type,Diamond(D)type,and Primitive(P)type were used.Through mathematical modeling of three-period minimal surface,different sizes and types of cells were used in the gradient region.A total of six kinds of cell gradient scaffolds(G-2P-4D,P-2D-4G,D-2P-4D,G-2D-4P,P-2G-4D,and D-2G-4P)were constructed and mechanical experiments and simulation experiments were conducted to evaluate the mechanical properties of the scaffolds.Flow performance parameters of the fluids in the scaffolds were obtained through computational fluid dynamics simulation.RESULTS AND CONCLUSION:Finite element mechanical simulation and compression experiment showed that P-2G-4D and P-2D-4G with the highest elastic modulus(148.67 MPa and 152.1 MPa),bearing a higher body load,improved the stability of the scaffold.The stress distribution in D-2P-4G was even and effectively reduced stress concentration,so that the connection function area could effectively transfer stress and reduce stress shielding.Flow rate was changing the least in G-2D-4P(0.10-0.48 mm/s).Permeability was higher than other scaffolds so that body fluids were able to flow though the gradient scaffold after implantation.This design method provides a new idea for the design of osteochondral scaffolds,and the simulation analysis results also provide a reference for the prediction of bone integration after implantation of scaffolds.

Objective:To investigate the impact of the number of examined lymph nodes (ELN) on survival outcomes in gastric cancer patients with postoperative pathological stage pN3b.Methods:This retrospective cohort study included 279 pN3b gastric cancer patients who underwent D2 gastrectomy at Nanfang Hospital, Southern Medical University (September 2008 to April 2023), with 35 patients receiving combination chemotherapy and anti-PD-1 therapy (immunotherapy group) and 244 receiving adjuvant chemotherapy alone (nonimmunotherapy group). Additionally, 422 patients with pN3b from the SEER database (2005 to 2020) were collected as an external validation cohort to determine the optimal cutoff value for the number of lymph nodes examined in the nonimmunotherapy group. The primary endpoints were overall survival (OS) and recurrence-free survival (RFS) in the nonimmunotherapy group of the Nanfang Hospital cohort, stratified by whether the number of examined lymph nodes was above or below the ELN optimal cutoff value. These findings were subsequently validated in the SEER cohort.Results:The optimal ELN cutoff value (34 nodes) was determined using X-tile software and by constructing an ELN-HR fitting model with inflection point identification. In the nonimmunotherapy group, patients with ELN >34 exhibited significantly prolonged survival compared to ELN ≤34 (median OS: 25.0 (95%CI:20.5-29.5) to 17.0 (95%CI:12.7-21.3) months, P=0.004; median RFS: 19.0 (95%CI:15.6-22.4) to 13.0 (95%CI:9.5-16.5) months, P=0.048). Multivariate Cox analysis also showed ELN >34 to be an independent protective factor for both OS (HR=0.576, 95%CI: 0.397-0.836) and RFS (HR=0.701, 95%CI: 0.492-0.998). In the SEER cohort, ELN >34 was associated with a 5-month OS extension (19 to 14 months, P=0.065), with multivariate analysis supporting its independent prognostic significance (HR=0.729, 95%CI: 0.580-0.915, P=0.006). Notably, in the immunotherapy group, patients with ELN >34 ( n=30) achieved a median OS of 41 months, but the median OS had not been reached in the ELN ≤34 group ( n=5) (1 death at 48 months). Conclusion:Higher ELN (>34) correlates with improved survival in nonimmunotherapy-treated pN3b gastric cancer patients. However, in pN3b gastric cancer patients treated with immunotherapy, the optimal ELN threshold requires further exploration to determine.

Objective:To construct a model of mitochondria-related genes (Mito-RGs) in hepatocellular carcinoma (HCC), and predict the immune, prognostic and therapeutic characteristics of HCC based on the model, so as to provide a new idea for the diagnosis and treatment of HCC.Methods:The expression profiles of HCC and corresponding clinical information were obtained from the Cancer Genome Atlas (TCGA) database. Univariate Cox regression, Least Absolute Shrinkage and Selection Operator (LASSO) regression and multivariate Cox regression were used to construct a prognostic model of HCC based on Mito-RGs, and the International Cancer Genome Consortium-Liver Cancer-RIKEN-Japan ICGC-LIRI-JP dataset were used for validation. GO and KEGG analyses were performed to investigate the signaling pathways enriched for differentially expressed genes in the high- and low-risk groups. Immune infiltration was assessed using CIBERSORT. Single-cell data were used to study the proportion of immune cells in high- and low-risk groups of HCC samples and the relationship with cell proliferation. Cellminer was used to assess the relationship between risk score models and drug sensitivity.Results:A risk-prognostic model of HCC containing seven Mito-RGs ( DTYMK, ACADS, HMGCS2, CYP27A1, TOMM40L, STOM, and AKR1B10) was constructed. High-risk HCC patients had a worse prognosis. Genes upregulated in the high- and low-risk groups of differentially expressed genes were enriched in cell cycle and metabolism-related pathways. Single-cell data showed higher proportions of CD8 + T cells, macrophages and monocytes, and proliferating cells in the high-risk group. CIBERSORT analysis suggested that Treg cells and M0 macrophages were more abundant in the high-risk group, whereas CD8 + T cells and CD4 + memory T cells were less abundant. Patients in the high-risk group were more sensitive to myeloid cell leukemia sequence 1 inhibitor, vincristine, phosphatidylinositol kinase beta subunit inhibitor, and aurora kinase A, while trametinib, selumetinib, extracellular regulated protein kinase, and mitogen-activated extracellular signal-regulated kinase were more effective in the low-risk group. Conclusion:The constructed Mito-RGs model is capable of providing a more accurate assessment of the prognosis and the degree of immune cell infiltration in HCC patients.