Objective: To fabricate a hydrogel loaded with inositol hexaphosphate-zinc and preliminarily evaluate its performance in self-mineralization and osteoinduction, thereby providing a theoretical basis for the development of bone regeneration materials. Methods: The hydrogel framework (designated DF0) was formed by copolymerizing methacryloyloxyethyltrimethylammonium chloride and four-armed poly(ethylene glycol) acrylate, followed by sequentially loading inositol hexaphosphate anions via electrostatic interaction and zinc ions via chelation. The hydrogel loaded only with inositol hexaphosphate anions was named DF1, while the co-loaded hydrogel was named DF2. The self-mineralization efficacy of the DF0 , DF1 and DF2 hydrogels was characterized using scanning electron microscopy, transmission electron microscopy (TEM), energy dispersive spectroscopy (EDS), and selected area electron diffraction (SAED). The biocompatibility was assessed via live/dead cell staining and a CCK-8 assay. The osteoinductive capacity of the DF0 , DF1 and DF2 hydrogels on MC3T3-E1 cells was assessed via alkaline phosphatase (ALP) and Alizarin Red S (ARS) staining. In the aforementioned cell experiments, cells cultured in standard medium served as the control group Results: The DF0, DF1, and DF2 hydrogels were successfully synthesized. Notably, DF1 and DF2 exhibited distinct self-mineralization within 6 days. Results from TEM, EDS, and SAED confirmed that the mineralization products were amorphous calcium phosphate in group DF1, and amorphous calciumzinc phosphate in group DF2. Biocompatibility tests revealed that none of the hydrogels (DF0, DF1, and DF2) adversely affected cell viability or proliferation. In osteogenic induction experiments, both ALP and ARS staining were intensified in the DF1 and DF2 groups, with the most profound staining observed in the DF2 group. Conclusion The developed inositol hexaphosphate-zinc hydrogel (DF2) demonstrates the dual capacity to generate calcium-phosphate compounds through self-mineralization while exhibiting excellent osteoinductive properties. This biocompatible, dual-promoting osteogenic hydrogel presents a novel strategy for bone regeneration.

Objective: To investigate the barrier membrane fixation performance and enhanced guided bone regeneration (GBR) capability of a thermosensitive adhesive containing bioactive glass nanoparticles in order to provide a novel solution for membrane fixation during GBR procedures. Methods: M2NP@BGN (methoxyethyl acrylate-co-N-isopropylacrylamide-co-protocatechuic acid@Bioactive glass nanoparticle), a thermosensitive adhesive, was synthesized via free radical polymerization by compositing methoxyethyl acrylate, N-isopropylacrylamide, and protocatechuic acid into a basic adhesive that was modified with bioactive glass nanoparticle (BGN). The successful fabrication of basic adhesive M2NP was characterized by attenuated total reflection-Fourier transform infrared spectroscopy and nuclear magnetic resonance spectroscopy. The thermosensitive adhesive M2NP@BGN (BGN concentration of 1 mg/mL) was characterized by scanning electron microscopy and a rheometer. By adjusting the BGN concentration (0.1 mg/mL, 0.5 mg/mL, 1 mg/mL, and 2 mg/mL), the adhesive and mechanical strengths were investigated with a universal testing machine. Biocompatibility was evaluated with a cell counting kit-8 assay and hemolysis test to identify the optimal formulation. The optimal material’s extract was co-cultured with mouse bone marrow mesenchymal stem cells, and its osteogenic activity was examined in vitro by quantitative real-time PCR, alkaline phosphatase, and alizarin red S staining. The rat mandibular defect model was established, filled with bone graft, and divided into 3 groups based on membrane fixation method: M2NP@BGN (BGN concentration of 1 mg/mL) fixation group (M2NP@BGN), titanium nail fixation group (Nail), and unfixed control group (Negative). Bone regeneration was analyzed after 8 weeks by micro computed tomography and histological staining. Results: M2NP@BGN (BGN concentration of 1 mg/mL) was successfully synthesized and demonstrated rapid gelation under warm, humid conditions. The adhesive with a BGN concentration of 1 mg/mL exhibited the highest adhesive strength (P < 0.001) and significantly enhanced mechanical strength (P < 0.001) under 37℃ wet conditions. All formulations showed excellent biocompatibility, with cell viability > 80% and hemolysis ratio < 5%. M2NP@BGN (BGN concentration of 1 mg/mL) significantly upregulated the expression of Runx2 and Col I (P < 0.001) and enhanced the activity of osteogenic differentiation markers (P < 0.05). In the animal model, the M2NP@BGN group (BGN concentration of 1 mg/mL) achieved significantly higher bone volume fraction and better bone maturity compared to the negative and nail groups (P < 0.05). Conclusion M2NP@BGN (BGN concentration of 1 mg/mL) combines excellent wet adhesion with potent osteogenic activity, enhances the bone augmentation efficacy of membranes, and presents a novel fixation strategy with significant clinical translation potential for GBR therapy.

Objective To explore the mechanism of GS-9620 in improving imiquimod(IMQ)-induced psoriasis-like inflammation by regulating the Th1/Th17-related immune response,and to investigate its regulatory effect on the gut microbiota in mice.Methods An IMQ-induced psoriasis-like inflammation model was established in BALB/c mice.The severity of the skin lesions was evaluated by psoriasis area and severity index(PASI)score.The proportions of CD4+interleukin(IL)-17+and CD4+interferon(IFN)-γ+cells in spleen tissue were detected by flow cytometry.Levels of the inflammatory factors tumor necrosis factor(TNF)-α,IL-1β,and IL-6 in skin tissues were determined by enzyme-linked immunosorbent assay,and pathological analysis was performed by hematoxylin/eosin staining.The effects of GS-9620 on the structure of the gut microbiota in control,IMQ model,and GS-9620-treated mice were detected by 16S rRNA sequencing.Results GS-9620 significantly reduced the PASI score in IMQ-induced mice and effectively reduced the proportions of CD4+IL-17+and CD4+IFN-γ+cells in the spleen.GS-9620 also significantly down-regulated the expression levels of TNF-α,IL-1β,and IL-6 in skin tissues.16S rRNA sequencing showed that GS-9620 significantly regulated the abundance of gut microbiota related to inflammation,including the relative abundances of bacteria such as Lachnospiraceae_NK4A136_group,Lachnospiraceae_UCG-008,Alloprevotella,Desulfovibrio,Prevotellaceae_UCG-001,and Alistipes.Conclusions GS-9620 effectively alleviates IMQ-induced psoriasis-like skin inflammation in mice by regulating the expression of Th1/Th17-related inflammatory factors.It may also improve IMQ-induced clinical symptoms by regulating the structure of the gut microbiota,thus providing a new theoretical basis for the treatment of psoriasis.The result of this study provide important experimental evidence to support further investigations into the application of GS-9620 for the treatment of psoriasis.

Objective To analyze the role and mechanism of PD-L1 in oral cancer metastasis based on TCGA and GEO databases.Methods The expression characteristics and clinical significance of the PD-L1 in oral cancer were analyzed using the TCGA database.PD-L1 mRNA levels were detected by quantitative reverse transcription-polymerase chain reaction(RT-qPCR)in various oral cancer cell lines.CCK-8,scrath test,Transwell-migration,and matrigel-invasion assays were employed to assess the effects of PD-L1 on proliferation,migration,and invasion of oral cancer cells.The interaction network between PD-L1 and functional genes in patients with oral cancer was constructed using STRING software and the GEO database,and key pathways were screened by Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.The regulatory relationship between PD-L1 and key genes was validated by RT-qPCR.Results TCGA data revealed that PD-L1 was highly expressed in patients with oral cancer and was correlated with lymph node metastasis(P＜0.01).PD-L1 was also highly expressed in oral cancer cell lines and its inhibition significantly inhibited the proliferation,migration,and invasion of Cal27 and SCC25 cells(P＜0.05).KEGG analysis indicated that PD-L1 activated the Janus kinase(JAK)/signal transducer and activator of transcription(STAT)pathway by upregulating C-X-C motif chemokine ligand(CXCL)9 and CXCL10,thereby promoting STAT1 expression to regulate oral cancer metastasis.Inhibition of the JAK/STAT pathway further suppressed the proliferation,migration,invasion,and expression of STAT1,CXCL9,and CXCL10 in Cal27 and SCC25 cells(P＜0.05).Conclusions PD-L1 may promote oral cancer cell proliferation,migration,and invasion by upregulating CXCL9 and CXCL10 to regulate the JAK/STAT pathway and enhance STAT1 expression,ultimately driving oral cancer growth and metastasis.

Objective To analyze the role and mechanism of PD-L1 in oral cancer metastasis based on TCGA and GEO databases.Methods The expression characteristics and clinical significance of the PD-L1 in oral cancer were analyzed using the TCGA database.PD-L1 mRNA levels were detected by quantitative reverse transcription-polymerase chain reaction(RT-qPCR)in various oral cancer cell lines.CCK-8,scrath test,Transwell-migration,and matrigel-invasion assays were employed to assess the effects of PD-L1 on proliferation,migration,and invasion of oral cancer cells.The interaction network between PD-L1 and functional genes in patients with oral cancer was constructed using STRING software and the GEO database,and key pathways were screened by Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.The regulatory relationship between PD-L1 and key genes was validated by RT-qPCR.Results TCGA data revealed that PD-L1 was highly expressed in patients with oral cancer and was correlated with lymph node metastasis(P＜0.01).PD-L1 was also highly expressed in oral cancer cell lines and its inhibition significantly inhibited the proliferation,migration,and invasion of Cal27 and SCC25 cells(P＜0.05).KEGG analysis indicated that PD-L1 activated the Janus kinase(JAK)/signal transducer and activator of transcription(STAT)pathway by upregulating C-X-C motif chemokine ligand(CXCL)9 and CXCL10,thereby promoting STAT1 expression to regulate oral cancer metastasis.Inhibition of the JAK/STAT pathway further suppressed the proliferation,migration,invasion,and expression of STAT1,CXCL9,and CXCL10 in Cal27 and SCC25 cells(P＜0.05).Conclusions PD-L1 may promote oral cancer cell proliferation,migration,and invasion by upregulating CXCL9 and CXCL10 to regulate the JAK/STAT pathway and enhance STAT1 expression,ultimately driving oral cancer growth and metastasis.

Objective To explore the mechanism of GS-9620 in improving imiquimod(IMQ)-induced psoriasis-like inflammation by regulating the Th1/Th17-related immune response,and to investigate its regulatory effect on the gut microbiota in mice.Methods An IMQ-induced psoriasis-like inflammation model was established in BALB/c mice.The severity of the skin lesions was evaluated by psoriasis area and severity index(PASI)score.The proportions of CD4+interleukin(IL)-17+and CD4+interferon(IFN)-γ+cells in spleen tissue were detected by flow cytometry.Levels of the inflammatory factors tumor necrosis factor(TNF)-α,IL-1β,and IL-6 in skin tissues were determined by enzyme-linked immunosorbent assay,and pathological analysis was performed by hematoxylin/eosin staining.The effects of GS-9620 on the structure of the gut microbiota in control,IMQ model,and GS-9620-treated mice were detected by 16S rRNA sequencing.Results GS-9620 significantly reduced the PASI score in IMQ-induced mice and effectively reduced the proportions of CD4+IL-17+and CD4+IFN-γ+cells in the spleen.GS-9620 also significantly down-regulated the expression levels of TNF-α,IL-1β,and IL-6 in skin tissues.16S rRNA sequencing showed that GS-9620 significantly regulated the abundance of gut microbiota related to inflammation,including the relative abundances of bacteria such as Lachnospiraceae_NK4A136_group,Lachnospiraceae_UCG-008,Alloprevotella,Desulfovibrio,Prevotellaceae_UCG-001,and Alistipes.Conclusions GS-9620 effectively alleviates IMQ-induced psoriasis-like skin inflammation in mice by regulating the expression of Th1/Th17-related inflammatory factors.It may also improve IMQ-induced clinical symptoms by regulating the structure of the gut microbiota,thus providing a new theoretical basis for the treatment of psoriasis.The result of this study provide important experimental evidence to support further investigations into the application of GS-9620 for the treatment of psoriasis.

Objectives:To explore the changes of some peripheral blood cells related to inflammation in patients with non-arteritis central retinal artery occlusion (NA-CRAO).Methods:A retrospective clinical study. From July 2019 to July 2021, a total of 218 patients with NA-CRAO hospitalized (NA-CRAO group) in Department of Ophthalmology, Xi'an People's Hospital (Xi'an Fourth Hospital) and 218 patients with routine physical examination (control group) during the same period were included in the study. There were no significant differences in age ( t=0.60), sex composition ratio ( χ2=0.83) and body mass index ( t=0.77) between the two groups ( P>0.05). 0.2 ml fasting peripheral blood was collected from the subject, and white blood cells (WBC), neutrophils (NEUT), lymphocytes (LYMPH), red blood cells (RBC), RBC distribution width (RDW), platelets (PLT), mean PLT volume (MPV), and large PLT ratio (PLCR) were detected. The NEUT/LYMPH ratio (NLR) and PLT/LYMPH ratio (PLR) were calculated. t test was used to compare measurement data between groups. Multiple logistic regression analysis was performed for blood cells with P <0.05. The receiver operating characteristic curve (ROC curve) was used to calculate the area under the curve (AUC) and 95% confidence interval (95% CI) of each inflammatory indicator, and the optimal cutoff value was determined according to the Jorden index (sensitivity+specificity-1). Results:Compared with control group, WBC, NEUT, NLR, RDW, PLR were increased in NA-CRAO group, while RBC and LYMPH were decreased, with statistical significance ( t=9.68, 12.43, 9.47, 3.64, 5.54, 5.18, 0.46; P<0.001). There was no significant difference in PLT, MPV and PLCR between the two groups ( t=0.32, 1.56, 0.84; P>0.05). Multivariate logistic regression analysis showed that NLR was a possible risk factor for the occurrence of NA-CRAO (odds ratio=2.51, 95% CI 0.780-0.859, P=0.031). ROC curve analysis showed that the AUC predicted by NLR was 0.819, the optimal critical value was 3.05, and the sensitivity and specificity were 59.2% and 92.7%, respectively. Conclusions:In peripheral blood cells of NA-CRAO patients, NEUT is significantly increased and LYMPH is decreased. NLR is a possible risk factor for NA-CRAO.

Objective:To observe the clinical and imaging features of non-arteriotic central retinal artery occlusion (NA-CRAO) with internal boundary membrane detachment (ILMD), and to analyze its relationship with visual prognosis.Methods:A retrospective clinical study. A total of 88 patients with NA-CRAO hospitalized in Department of Ophtalmology, Xi'an People's Hospital (Xi'an Fourth Hospital) from January 2014 to June 2023 were included in the study. Best corrected visual acuity (BCVA), optical coherence tomography (OCT) and fluorescein fundus angiography (FFA) were performed. The BCVA test used the international standard visual acuity chart, which was statistically converted to the logarithm of the minimum angle of resolution (logMAR) visual acuity. OCT observed the presence of ILMD and the thickening of the inner retina and the disappearance of anatomical stratification. FFA recorded arm-retinal circulation time (A-Rct) and retinal arterion-distal filling time (FT), and observed ciliary retinal artery, fluorescein retrograde filling, cotton spots, luciferin nodal filling, macular non-perfusion, capillary fluorescein leakage, optic disc strong fluorescence, choroidal background weak fluorescence and other characteristics. According to whether there was ILMD, the patients were divided into ILMD group and non-ILMD group, with 44 cases and 44 eyes respectively. The two groups received the same treatment. The follow-up time was 30 days after treatment. The clinical, FFA characteristics and BCVA before and after treatment were compared between the two groups. t-test was used for comparison between groups. Results:In ILMD group and non-ILMD group, there were 43 cases of male and 1 case of female, respectively, and the proportion of male was significantly higher than that of female. Before and after treatment, the logMAR BCVA of ILMD group and non-ILMD group were 2.35±0.42, 2.01±0.46, 1.47±0.60, 0.77±0.49, respectively. There were significant differences in logMAR BCVA between the two groups before and after treatment ( t=8.025, 12.358; P<0.001). Before treatment, A-Rct and FT in ILMD group were longer than those in non-ILMD group, and the difference was statistically significant ( t=3.052, 3.385; P<0.05). After treatment, there was no significant difference ( t=1.040, 1.447; P >0.05). The proportion of ciliary retinal artery and cotton plaque in ILMD group was lower than that in non-ILMD group. There was no significant difference in ciliary retinal artery between the two groups ( χ2=-0.961, P>0.05), but there was a significant difference in cotton wool plaque between the two groups ( χ2=-3.364, P <0.05). Compared to the non-ILMD group, The proportion of retrograde fluorescein filling in retinal artery ( χ2=-2.846), segment filling ( χ2=-3.907), macular non-perfusion ( χ2=-6.656), capillary fluorescein leakage ( χ2=-4.367), optic disc strong fluorescence ( χ2=-3.525) and choroidal background weak fluorescence ( χ2=-2.276) increased, the difference was statistically significant ( P<0.05). Conclusions:In patients with NA-CRAO, compared with those without ILMD, those with ILMD have more severe retinal ischemia and worse BCVA before and after treatment. ILMD is one of the poor prognostic markers of NA-CRAO vision.

Objective To investigate the effects of Gli2 on the proliferation,growth,migration,and invasion of oral cancer cells(Tca8113)at the cellular level,and to clarify the molecular mechanism of how Gli2 regulation affects the migration and invasion of oral cancer cells.Methods Small interfering(si)RNA was used to inhibit Gli2 expression in Tca8113 cells.The effects of Gli2 on the proliferation,growth,migration,and invasion of Tca8113 cells were examined by CCK-8,platb cloning,and transwell chamber assay.Further qRT-PCR and Western blot assays were used to explore the mechanism of how Gli2 regulation effects the malignant proliferation and metastasis of Tca8113 cells.Results The mRNA and protein expression of Gli2 in oral cancer cells(Tca8113)increased.Interference of Gli2 expression inhibited the proliferation,growth,migration,and invasion of Tca8113 cells.Further experiments showed that interfering with Gli2 expression inhibited the mRNA and protein expression of key factors in the Hedgehog(Hh)pathway.In addition,interference of Gli2 expression significantly affected the mRNA and protein expression of key factors in epithelial mesenchymal transformation(EMT)pathways.Conclusions Gli2 is abnormally activated during oral cancer,and interference of Gli2 expression significantly inhibits the proliferation,growth,migration,and invasion of oral cancer cells.Gli2 influences the migration and invasion of oral cancer cells by regulating the Hh and EMT pathways.This study has provided a new way to elucidate the pathogenesis of oral cancer and new perspectives on the clinical treatment of oral cancer.

Immunodeficient animal models play an important role in preclinical research and are important experimental tools in modern biomedical research that are widely used in immunology,genetics,oncology,microbiology,and other research fields.Gene editing is a technology for targeted modification of biological genomes.From emergence to application,it has greatly promoted the development of biomedical research.Gene editing technology mainly includes homing endonucleases,zinc finger nucleases,transcription activator-like effector nucleases,and the CRISPR/Cas9 system.Researchers have used these technologies to establish various types of immunodeficient animal models,each with advantages and limitations.In recent years,a large number of studies have confirmed that the human immunodeficient animal model accurately simulates the functions of cancer cells,drugs,and the human immune system,better simulates human diseases,and is widely used to study human immunobiology and the potential mechanisms of complex diseases.In this article,we review the progress in the research and application of gene editing technology to the establishment of immunodeficient animal models,discuss in depth the problems and optimization strategies of gene editing technology in the preparation of immunodeficient animal models,and present its future development prospects to provide references for researchers to select and establish immunodeficient animal models.