Objective To systematically evaluate the similarities between six-gene-modified pig red blood cells and human red blood cells in multiple dimensions, including cell morphology, hematological parameters, microbial safety and serological compatibility, and to verify the feasibility of using six-gene-modified pig red blood cells as an alternative for blood transfusion in cases of traumatic hemorrhage. Methods Blood samples from O-type human donor, six-gene-modified O-type pig and wild-type O-type pig were collected. The pig gene-modified phenotype was identified by flow cytometry. The morphology was observed using field emission scanning electron microscopy and the diameter was measured. Hematological tests, including blood routine, osmotic fragility test, adenosine triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) content and partial pressure of oxygen at 50% hemoglobin saturation (P50), were conducted to assess the deformability of red blood cells. Real-time fluorescent quantitative polymerase chain reaction was used to screen for zoonotic viruses, and cross-matching was performed using column gel method, condensation amine method and saline tube method. Results The six-gene-modified pig red blood cells successfully knocked out the α-Gal, Neu5Gc and Sda antigens and expressed hCD55. Wild-type pig red blood cells expressed these three antigens but did not express human complement regulatory proteins. All three types of red blood cells presented a biconcave discoid morphology. The diameter of O-type human red blood cells was larger than that of the six-gene-modified porcine red blood cells and wild-type pig red blood cells (all P < 0.05). The average red blood cell volume and average red blood cell hemoglobin of O-type humans were higher than those of the six-gene-modified pig red blood cells, and the distribution width-variable coefficient of O-type human red blood cells was lower than that of the six-gene-modified pig red blood cells (all P < 0.05). There were no statistically significant differences in the permeability fragility, ATP content, 2,3-DPG content, P50 and red blood cell deformability indicators of the three types of red blood cells (all P > 0.05). The screening for zoonotic viruses in human and animal cells of the six-gene-modified pigs and wild-type pigs was negative. The cross-matching results showed that in the column gel method and condensation amine method, the primary and secondary sides of the six-gene-modified pig red blood cells did not agglutinate with human red blood cells, while the wild-type pig red blood cells agglutinated with human red blood cells in all cross-matching methods. Conclusions The six-gene-modified pig red blood cells are highly similar to human red blood cells in terms of morphology, key hematological parameters, energy metabolism, oxygen-carrying capacity, mechanical properties, microbial safety and serological compatibility, providing an experimental basis for the clinical transformation of xenotransfusion.

Objective:To establish a prognostic model based on endoplasmic reticulum stress-related genes for evaluating the prognosis of patients with renal cell carcinoma.Methods:This study utilized Non-negative Matrix Factorization to identify molecular subgroups based on endoplasmic reticulum stress-related genes and employed Weighted Correlation Network Analysis to determine co-expressed genes associ-ated with these subgroups.A risk prognostic model was constructed using univariate Cox regression analysis and Lasso regression analysis.Preliminary experimental validations were conducted to elucidate the biological functions of model genes in renal cell carcinoma.Results:Two molecular subgroups with distinct survival prognoses were identified,and an intersection of related genes was used to construct a nov-el endoplasmic reticulum stress-related prognostic model.Patients in the high-risk group exhibited significantly poorer overall survival in both the training and validation cohorts.In vivo experiments demonstrated that PCK1,a model gene,could inhibit the proliferation,migra-tion,and invasion of renal cell carcinoma cells.Conclusions:The risk scoring model developed in this study effectively predicts the survival probability of renal cell carcinoma patients and can serve as an independent prognostic indicator.This model offers a new direction for per-sonalized treatment strategies in renal cell carcinoma patients.

ObjectiveTo investigate the possible mechanism by which Zhiqiao Gancao Decoction （枳壳甘草汤， ZGD） delays intervertebral disc degeneration （IDD） based on the Hippo-yes-associated protein （YAP）/transcriptional co-activator with PDZ-binding motif （TAZ） signaling pathway. MethodsA total of 50 SD rats were randomly divided into sham surgery group， model group， low-dose ZGD group， high-dose ZGD group， and high-dose ZGD + inhibitor group， with 10 rats in each group. In the sham surgery group， the rats were pierced in the skin and muscle at the Co6/7/8 segments of the tail with a 21G needle （depth approximately 2 mm） without damaging the intervertebral disc. In the other groups， rats were injected with a 21G needle at the Co6/7/8 segments of the tail to establish an IDD model by piercing the tail intervertebral disc 5 mm. One week after modeling， rats in the low-dose and high-dose ZGD groups were given 6.24 and 12.24 g/（kg·d） of the decoction via gastric gavage， respectively. The high-dose ZGD + inhibitor group was given 12.24 g/（kg·d） of the decoction and an intraperitoneal injection of YAP/TAZ inhibitor Verteporfin 10 mg/kg. The sham surgery and model groups were given 5 ml/（kg·d） of normal saline via gavage. The gavage was given once a day， and the intraperitoneal injection was given every other day. After 4 weeks of continuous intervention， the pathological changes of the tail intervertebral discs were observed using HE staining， Oil Red O-Green staining， and Toluidine Blue staining. Immunohistochemistry was used to detect the expression of aggrecan and MMP3 in the nucleus pulposus. TUNEL fluorescence staining was performed to detect apoptosis in the nucleus pulposus， and the apoptosis rate was calculated. Western blot was used to detect the Hippo-YAP/TAZ signaling pathway， including YAP， phosphorylated YAP （p-YAP）， phosphorylated MST1/2 （p-MST1/2）， phosphorylated TAZ （p-TAZ） and apoptosis-related proteins， such as Cleaved Caspase 3， P53， Bcl-2 and Bax. ResultsCompared with sham surgery group， the rats in the model group showed significant degenerative changes in the intervertebral disc. The levels of aggrecan， Bcl-2， and YAP proteins in the nucleus pulposus decreased， while the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and the apoptosis rate increased （P < 0.01）. Compared with the model group， the drug intervention groups showed partial recovery in intervertebral disc degeneration. The levels of aggrecan， Bcl-2， and YAP proteins increased， while the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and the apoptosis rate decreased （P＜0.05 or P＜0.01）. The high-dose ZGD group showed more significant recovery in intervertebral disc degeneration compared to the low-dose ZGD group， with a decrease in the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and apoptosis rate， and an increase in the levels of aggrecan， Bcl-2， and YAP proteins （P＜0.05 or P＜0.01）. Compared with the high-dose ZGD group， the high-dose ZGD + inhibitor group showed a reduced recovery in intervertebral disc degeneration， with an increase in the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and apoptosis rate， and a decrease in the levels of aggrecan， Bcl-2， and YAP proteins （P＜0.05 or P＜0.01）. ConclusionZGD may delay intervertebral disc degeneration by inhibiting the phosphorylation of YAP in the nucleus pulposus， maintaining the function of the Hippo-YAP/TAZ signaling pathway， and reducing apoptosis of nucleus pulposus cells.

OBJECTIVE To explore the effect and mechanism of Zhiqiao gancao decoction （ZQGCD） on pathological angiogenesis of degenerative intervertebral disc. METHODS The rats were randomly divided into sham operation group （normal saline）， model group （normal saline）， hypoxia inducible factor-1α （HIF-1α） inhibitor （YC-1） group [2 mg/（kg·d）， tail vein injection]， and ZQGCD low-dose， medium-dose and high-dose groups [3.06， 6.12， 12.24 g/（kg·d）]， with 8 rats in each group. Except for sham operation group， lumbar disc degeneration model of rat was constructed in all other groups. After modeling， they were given relevant medicine once a day， for consecutive 3 weeks. After the last medication， pathological changes and angiogenesis of the intervertebral disc tissue in rats were observed； the levels of inflammatory factors [interleukin-1β （IL-1β）， IL-6， tumor necrosis factor-α （TNF-α）] and the expressions of angiogenesis-related proteins [HIF-1α， vascular endothelial growth factor （VEGF）， VEGF receptor 2 （VEGFR2）， angiotensin 1（Ang 1）， Ang 2] in the com intervertebral disc tissue in rats were all determined. In cell experiment， the primary nucleus pulposus cells were isolated and cultured from rats， and cellular degeneration was induced using 50 ng/mL TNF-α. The cells were divided into blank control group （10% blank control serum）， TNF-α group （10% blank control serum）， YC-1 group （10% blank control serum+0.2 mmol/L YC-1）， and 5%， 10%， 15% drug-containing serum group （5%， 10%， 15% drug-containing serum）. After 24 hours of intervention， the nucleus pulposus cells were co-cultured with HUVEC. The expressions of Collagen Ⅱ， matrix metalloproteinase-3 （MMP-3） in nucleus pulposus cells were detected. HUVEC proliferation， migration and tube forming ability were detected， and the expression levels of the HIF-1α/VEGF/Ang signal axis and angiogenesis- related proteins （add MMP-2， MMP-9） in HUVEC were detected. RESULTS Animal experiments had shown that compared with model group， the positive expression of CD31 in the intervertebral disc tissues of rats in each drug group was down-regulated （P＜ 0.05）， the levels of inflammatory factors and angiogenesis-related proteins were decreased significantly （P＜0.05）， and the pathological changes in the intervertebral disc were alleviated. Cell experiments had shown that compared with TNF-α group， the expression of Collagen Ⅱ in nucleus pulposus cells of all drug groups was significantly up-regulated （P＜0.05）， and the expression of MMP-3 was significantly down-regulated （P＜0.05）； the proliferation， migration and tubulogenesis of HUVEC were significantly weakened （P＜0.05）. The mRNA and protein expressions of HIF-1α， VEGF， Ang 2 as well as the expression of angiogenesis-related proteins （except for the expression of Ang 2 mRNA and HIF-1α， VEGFR2， Ang 2 protein in 5% drug- containing serum group） were significantly down-regulated （P＜0.05）. CONCLUSIONS ZQGCD may inhibit the HIF-1α/VEGF/ Ang signal axis to weaken the angiogenic ability of vascular endothelial cells， improve pathological angiogenesis in the intervertebral disc， and delay the degeneration of the intervertebral disc.

IL-2 is a multifunctional cytokine whose main functions include activating and maintaining T cell survival.Re-search has shown that the level and function of IL-2 in peripheral blood of patients with systemic lupus erythematosus(SLE)have sig-nificantly decreased,and are associated with the activity of SLE disease.In recent years,the decrease in IL-2 level has been found to play a crucial role in the occurrence and development of SLE,and is one of the important mechanisms for T cell homeostasis imba-lance and immune regulation disorders in SLE patients.The low-dose IL-2 treatment regimen for SLE based on this discovery has en-tered clinical application.However,there is still considerable controversy regarding the mechanism of decreased IL-2 level and sup-pressed functions in SLE patients.Clarifying this mechanism is of great significance for further clarifying the occurrence and develop-ment mechanism of SLE and developing new treatment regimens.

Objective To investigate the clinical value of 3.0 T functional magnetic resonance imaging in evaluating postoperative recurrence and treatment efficacy of glioma. Methods A retrospective analysis was conducted on the general clinical data of 67 patients who underwent glioma surgery at the Second Affiliated Hospital of Xingtai Medical University. All patients received chemotherapy for more than one month post-surgery. Recurrence of glioma was diagnosed based on secondary surgery or pathological biopsy results as the gold standard. From 3 to 6 months post-surgery, computerized tomography was used to measure cerebral blood volume (CBV), three-dimensional arterial spin labeling was used to measure cerebral blood flow (CBF) and relative CBF (rCBF), and diffusion-weighted imaging with and without contrast enhancement was used to measure apparent diffusion coefficient (ADC). Data were analyzed using SPSS 26.0 statistical software, and the t test or χ² test was used for inter-group comparisons based on data type. The receiver operating characteristic (ROC) curve was applied to evaluate the value of CBV, rCBF, and ADC in assessing postoperative recurrence and treatment efficacy of glioma. Results Patients with high-grade gliomas showed significantly higher CBV and rCBF and significantly lower ADC compared to those with low-grade gliomas (P < 0.05). The area under the ROC curve (AUC) of CBV, rCBF, and ADC in combination for grading glioma was 0.960, which was higher than those of individual indicators (0.790, 0.955, and 0.795, P < 0.05). The recurrence group had significantly higher CBV and rCBF and lower ADC compared to the non-recurrence group (P < 0.05). The AUC of CBV, rCBF, and ADC in combination for predicting postoperative glioma recurrence was 0.965, which was significantly higher than those of individual indicators (0.729, 0.929, and 0.941, P < 0.05). CBV and rCBF were lower and ADC was higher in the effective treatment group than in the ineffective treatment group (P < 0.05). The AUC of CBV, rCBF, and ADC in combination for evaluating glioma treatment efficacy was 0.985, which was higher than those of individual indicators (0.842, 0.898, and 0.961, P < 0.05). Conclusion The CBV, rCBF, and ADC in combination has shown high diagnostic accuracy and predictive efficacy in the evaluation of postoperative recurrence and treatment efficacy of glioma, which has important clinical application value.

Objective:To establish a prognostic model based on endoplasmic reticulum stress-related genes for evaluating the prognosis of patients with renal cell carcinoma.Methods:This study utilized Non-negative Matrix Factorization to identify molecular subgroups based on endoplasmic reticulum stress-related genes and employed Weighted Correlation Network Analysis to determine co-expressed genes associ-ated with these subgroups.A risk prognostic model was constructed using univariate Cox regression analysis and Lasso regression analysis.Preliminary experimental validations were conducted to elucidate the biological functions of model genes in renal cell carcinoma.Results:Two molecular subgroups with distinct survival prognoses were identified,and an intersection of related genes was used to construct a nov-el endoplasmic reticulum stress-related prognostic model.Patients in the high-risk group exhibited significantly poorer overall survival in both the training and validation cohorts.In vivo experiments demonstrated that PCK1,a model gene,could inhibit the proliferation,migra-tion,and invasion of renal cell carcinoma cells.Conclusions:The risk scoring model developed in this study effectively predicts the survival probability of renal cell carcinoma patients and can serve as an independent prognostic indicator.This model offers a new direction for per-sonalized treatment strategies in renal cell carcinoma patients.

Objective To explore the function and molecular mechanisms of matrix metalloproteinase 19(MMP-19)in colorectal cancer(CRC)and liver metastasis of CRC(CRLM).Methods This study comprehen-sively analyzed transcriptomic data,single-cell data,and clinical information of CRC and CRLM patients retrieved from public databases,namely The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO).RNA sequencing analysis was carried out to identify biological functions associated with MMP-19 expression.The differ-ential expression of MMP-19 between primary lesions and liver metastatic lesions was evaluated using quantitative real-time polymerase chain reaction(qRT-PCR),Western blotting,and immunohistochemistry.In vitro functional assays were performed to validate the impact of MMP-19 on the proliferation,invasion,and migration of colorectal cancer cells.Results MMP-19 was highly expressed in CRC tissues and was significantly correlated with poor prognosis in both CRC and CRLM patients.Sequencing results and functional enrichment analysis demonstrated that MMP-19 is involved in epithelial-mesenchymal transition,amino acid and protein transport,cell proliferation,extracellular matrix organization,and the mitogen-activated protein kinase(MAPK)signaling pathway.Single-cell analysis revealed that the expression of MMP-19 in macrophages and dendritic cells was higher in both CRC primary tumors and liver metastases compared to normal tissues.In vitro experiments confirmed that MMP-19 enhances the proliferation,invasion,and migration capabilities of colorectal cancer cells.Conclusion MMP-19 has been shown to play a crucial role in promoting the development and progression of CRC.It increases the risk of postoperative recurrence and liver metastasis through inducing epithelial-mesenchymal transition and remodeling the immune microenvironment.Given these findings,MMP-19 emerges as a promising novel target for the diagnosis and treatment of both CRC and CRLM.

Objective To explore the function and molecular mechanisms of matrix metalloproteinase 19(MMP-19)in colorectal cancer(CRC)and liver metastasis of CRC(CRLM).Methods This study comprehen-sively analyzed transcriptomic data,single-cell data,and clinical information of CRC and CRLM patients retrieved from public databases,namely The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO).RNA sequencing analysis was carried out to identify biological functions associated with MMP-19 expression.The differ-ential expression of MMP-19 between primary lesions and liver metastatic lesions was evaluated using quantitative real-time polymerase chain reaction(qRT-PCR),Western blotting,and immunohistochemistry.In vitro functional assays were performed to validate the impact of MMP-19 on the proliferation,invasion,and migration of colorectal cancer cells.Results MMP-19 was highly expressed in CRC tissues and was significantly correlated with poor prognosis in both CRC and CRLM patients.Sequencing results and functional enrichment analysis demonstrated that MMP-19 is involved in epithelial-mesenchymal transition,amino acid and protein transport,cell proliferation,extracellular matrix organization,and the mitogen-activated protein kinase(MAPK)signaling pathway.Single-cell analysis revealed that the expression of MMP-19 in macrophages and dendritic cells was higher in both CRC primary tumors and liver metastases compared to normal tissues.In vitro experiments confirmed that MMP-19 enhances the proliferation,invasion,and migration capabilities of colorectal cancer cells.Conclusion MMP-19 has been shown to play a crucial role in promoting the development and progression of CRC.It increases the risk of postoperative recurrence and liver metastasis through inducing epithelial-mesenchymal transition and remodeling the immune microenvironment.Given these findings,MMP-19 emerges as a promising novel target for the diagnosis and treatment of both CRC and CRLM.

IL-2 is a multifunctional cytokine whose main functions include activating and maintaining T cell survival.Re-search has shown that the level and function of IL-2 in peripheral blood of patients with systemic lupus erythematosus(SLE)have sig-nificantly decreased,and are associated with the activity of SLE disease.In recent years,the decrease in IL-2 level has been found to play a crucial role in the occurrence and development of SLE,and is one of the important mechanisms for T cell homeostasis imba-lance and immune regulation disorders in SLE patients.The low-dose IL-2 treatment regimen for SLE based on this discovery has en-tered clinical application.However,there is still considerable controversy regarding the mechanism of decreased IL-2 level and sup-pressed functions in SLE patients.Clarifying this mechanism is of great significance for further clarifying the occurrence and develop-ment mechanism of SLE and developing new treatment regimens.