Type 2 diabetes (T2D) is an independent risk factor for cognitive impairment. The dysregulation of hypoxia inducible factor (HIF) signaling in T2D patients results in impaired adaptive responses to hypoxia, thereby accelerating the progression of complications. However, limited knowledge is available regarding its precise function in diabetes-associated cognitive impairment (DACI). Here, elevated HIF-1α levels were observed in brain endothelial cells (ECs) of db/db mice. Functionally, brain ECs-specific knockdown of H if1 a significantly ameliorated T2D-induced memory loss and neuronal damage. Glycolysis in brain ECs was inhibited in this process, as indicated by RNA-seq, leading to decreased hippocampal lactate production through reduced LDHA expression. Notably, T2D patients showed increased cerebrospinal fluid lactate levels, which were strongly associated with their cognitive dysfunction. Intrahippocampal injection of lactate accelerated cognitive dysfunction and impaired adult hippocampal neurogenesis (AHN) in db/db mice. Conversely, reducing hippocampal lactate levels through the intrahippocampal injection of oxamate delayed the onset of memory deficits. Furthermore, asiatic acid was discovered to protect db/db mice from cognitive impairment by decreasing brain endothelial HIF-1α expression and subsequently reducing hippocampal lactate-induced AHN damage. Overall, this study elucidates the inhibiting role played by endothelial HIF-1α-driven lactate in AHN and highlights a potential tactic of targeting HIF-1α in brain ECs for treating cognitive impairment.

BACKGROUND:Bile acid metabolism plays a crucial role in the development and progression of Crohn's disease.There is no research on changes in bile acid metabolism and key target genes following treatment with biological agents.OBJECTIVE:To investigate the expression characteristics of bile acid metabolism-related genes in patients with Crohn's disease,identify key genes associated with response to biological agents.METHODS:Transcriptome data were obtained through the GEO database to analyze differentially expressed genes between inflammation-control groups and inflammation-treatment groups.GO,KEGG,and GSEA enrichment analyses were used to evaluate the effects of biological agent therapy on bile acid metabolism.Protein-protein interaction network and WGCNA algorithm were employed to analyze differentially expressed genes,identifying modules closely related to biological agent treatment response,which led to the determination of UGT2A3 as a key gene in bile acid metabolism.In the inflammation group of the GSE186582 dataset,samples were divided into high and low expression groups based on UGT2A3 levels to study its relationship with immune infiltration and explore the interaction between UGT2A3 and the immune microenvironment.Clinical characteristics and intestinal manifestations were compared between high and low expression groups,and correlations between UGT2A3 and clinical indicators(C-reactive protein,erythrocyte sedimentation rate,Crohn's disease activity index,and Crohn's disease endoscopic activity score)were investigated.The competing endogenous RNA regulatory network of UGT2A3 was constructed,and its upstream miRNA was functionally enriched to explore the molecular mechanism of UGT2A3 in bile acid metabolism.Single-cell analysis and clustering were performed using high-throughput sequencing data of GSE134809 to observe the expression of UGT2A3 in different samples and cell populations.Colon tissue samples from untreated and biologic-treated Crohn's disease patients and healthy colon tissue samples from patients with intestinal polyps were collected,and UGT2A3 expression was detected by immunohistochemistry,qRT-PCR,and western blot assay.Fresh feces from Crohn's disease patients and healthy controls were collected to detect bile acid levels,and the relationship between UGT2A3 and fecal bile acid levels was analyzed.RESULTS AND CONCLUSION:A total of 11 bile acid metabolism-related genes were screened,showing significant changes in gene expression after biological agent therapy.GO and KEGG enrichment analyses revealed that intestinal nutrient absorption and metabolic processes normalized after treatment,while leukocyte chemotaxis and inflammatory response pathway activity decreased.GSEA analysis revealed significant enrichment of bile acid metabolism-related pathways after treatment.Protein-protein interaction network construction and WGCNA analysis identified UGT2A3 as a key gene closely associated with treatment response.UGT2A3 expression was significantly decreased in inflamed tissues of Crohn's disease patients and returned to normal levels after biological agent therapy.This result was confirmed in clinical specimens.UGT2A3 expression levels showed significant negative correlations with C-reactive protein,erythrocyte sedimentation rate,Crohn's Disease Activity Index,and Crohn's Disease Endoscopic Index of Severity.Receiver Operating Characteristic curve analysis demonstrated that UGT2A3 has good diagnostic value(Area Under Curve AUC=0.801 0)and effectively reflects treatment outcomes.Immune infiltration analysis showed significantly increased infiltration of various immune cells in samples with low UGT2A3 expression,and its expression levels negatively correlated with immune scores,microenvironment scores,and stromal scores.Compared with the low UGT2A3 expression group,patients with high expression showed less fecal occult blood and penetrating inflammation,with milder intestinal strictures and general condition severity.Fecal bile acid analysis revealed that UGT2A3 expression strongly negatively correlated with primary bile acid content and strongly positively correlated with secondary bile acid content.

Objectives:To investigate the expression of signal-induced proliferation-associated 1 (SIPA1) in colorectal cancer tissues and its relationship with patient prognosis. To explore the effects of SIPA1 on proliferation and migration abilities, as well as the possible molecular mechanisms.Methods:Using The Cancer Genome Atlas (TCGA) database to analyze the differential expression of SIPA1 and conduct survival analysis. Then, plotting receiver operating characteristic curve (ROC) and prognosis calibration curve analysis to assess the predictive capability and accuracy of SIPA1 for patient prognosis. Subsequently, verifying the expression levels of SIPA1 in tumor tissues and adjacent normal tissues using immunohistochemistry (IHC) and western blot (WB) assays(from March 1, 2023, to May 1, 2024, pathological specimens of five colorectal cancer patients were selected from the tissue bank of affiliated hospital of Nantong University. tissue microarrays were constructed using both cancerous tissues and adjacent normal tissues), and exploring the correlation between SIPA1 and clinical pathological parameters. Next, establishing SIPA1 stable knockdown cell lines in colorectal cancer cell lines DLD1 and HCT116, and assessing the biological behavior changes of tumor cells after SIPA1 knockdown through cell proliferation, invasion, and migration experiments. Validating the impact of SIPA1 on colorectal cancer cell proliferation in vivo through subcutaneous xenograft experiments in nude mice. Exploring the potential pro-tumor mechanisms of SIPA1 through pathway enrichment analysis, and confirming these using WB experiments. The proliferation, invasion and migration of tumor cells were detected after adding PI3K activator. Lastly, conducting correlation analysis between SIPA1 and immune checkpoint, as well as the association with immune cells in the tumor microenvironment. Results:Differential analysis showed that mRNA expression of SIPA1 in colorectal cancer tissues was significantly higher than that in adjacent normal tissues ( P＜0.05). Prognostic analysis indicated that patients with high expression of SIPA1 had poor overall survival ( P＜0.001), and the expression level of SIPA1was correlated with lymph node invasion ( P＜0.001) and N stage ( P＜0.05). ROC curve and prognosis calibration curve suggest that SIPA1 can effectively predict five-year survival rate of patients (AUC=0.7), and the predictive performance of the model is relatively accurate ( P＜0.001). WB experiments showed a significant increase in the expression level of SIPA1 protein in colorectal cancer specimens ( P＜0.001). Immunohistochemistry results indicated higher staining scores of SIPA1 in tumor tissues. In vitro experiments demonstrated that SIPA1 knockdown significantly inhibited the proliferation, invasion, and migration capabilities of colorectal cancer cells. In DLD1 and HCT116 cells, the SIPA1-knockdown group exhibited significantly lower absorbance compared to the control group (0.89±0.01 vs. 1.57±0.02 and 0.72±0.01 vs. 1.31±0.03, respectively, both P＜0.001). The SIPA1-knockdown group also demonstrated a reduced number of migrated cells relative to the control group (197.93±16.64 vs. 518.48±29.15 and 171.83±12.49 vs. 446.00±21.81, respectively, both P＜0.001). Furthermore, the cell wound-healing rate was significantly lower in the SIPA1-knockdown group than that in the control group [(0.32±0.01)% vs. (0.61±0.01)% and (0.28±0.01)% vs. (0.75±0.01)%, respectively, both P＜0.001]. In vivo animal experiments suggested that SIPA1 knockdown could inhibit tumor growth [(460.35±57.47) mm3 vs (1 177.55±208.24)mm3, (0.76±0.11)g vs (1.43±0.08)g, P＜0.05]. Pathway enrichment analysis revealed significant enrichment of the receptor tyrosine kinase signaling pathway, and SIPA1 knockdown could inhibit the activation of the phosphatidylinositide 3-kinases (PI3K)/protein kinase B (PKB)/glycogen synthase kinase-3β (GSK3β) signaling pathway. The PI3K activator reversed the inhibitory effect of SIPA1 silencing on tumor cell proliferation, invasion and migration. Correlation analysis indicated that high expression of SIPA1 was associated with immune checkpoints and various immunosuppressive cells (all P＜0.05). Conclusions:SIPA1 is highly expressed in colorectal cancer and associated with poor prognosis. SIPA1 may affect the proliferation and migration abilities of tumor cells by regulating the PI3K/AKT/GSK3β signaling pathway.

Objective:To investigate the relationship between serum apolipoprotein A1 (ApoA1), cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), proinflammatory protein S100A9 (S100A9) and prognosis in patients with myelodysplastic syndrome (MDS).Methods:122 MDS patients visited Qinhuangdao First Hospital from January 2020 to January 2021 were selected as the research objects. After three years of follow up, patients were divided into survival group and death group based on survival status. The differences in ApoA1, CTLA-4, and S100A9 between the death group and the survival group were compared. Measurement data with normal distribution was expressed as " xˉ±s", independent sample t-test was used on comparison between groups. Counting data was expressed as rate or composition ratio, χ2 test was used on comparison between groups. Univariate and multivariate Logistic regression analysis were used to analyze factors related to death. Results:There was no lost to follow up patients after three years of follow up. Among those 122 patients, 92 survived and 30 died. The ratio of bone marrow primitive cells>5%, IPSS-R score, serum CTLA-4, and S100A9 levels in the survival group were (2.89±2.15), (3.13±1.95) points, (5.12±1.59) μg/L, (1643.98±429.65)ng/L, respectively, lower than (5.67±3.76), (5.12±2.36) points, (28.67±6.98) μg/L, (2895.64±553.62) ng/L in the death group ( t=5.03, 4.60, 30.27, 12.87, respectively, all P<0.01). The relative high-risk ratio of IPSS-R stratification in the survival group was 63.04%,(58/92) which was lower than the 86.67%(26/30) in the death group ( χ2=5.89, P=0.015). The absolute values of hemoglobin, lymphocytes and neutrophils, and values of platelets and ApoA1 in the survival group were(86.74±12.69)g/L, (1.41±0.23)×10 9/L, (1.42±0.55)×10 9/L, (59.98±21.37)×10 9/L, (1.09±0.40) g/L respectively, which were higher than (65.58±10.89)g/L, (0.68±0.17)×10 9/L, (0.96±0.31)×10 9/L, (42.85±20.95)×10 9/L, (0.91±0.36)g/L in the death group ( t=8.20, 16.00, 4.35, 7.90, 2.19; respectively, P<0.001, <0.001, <0.001, <0.001, =0.030). Multivariate Logistic regression model analysis showed that, bone marrow blasts cells>5% ( OR=1.732, 95% CI: 1.188~2.523, P=0.004), relatively high IPSS-R stratification ( OR=1.815, 95% CI: 1.332~2.474, P<0.001), high IPSS-R score ( OR=1.785, 95% CI: 1.259~2.529, P=0.001), high CTLA-4 level ( OR=2.156, 95% CI: 1.482~3.134, P<0.001) and high S100A9 level ( OR=1.787, 95% CI: 1.218~2.625, P=0.003) were risk factors for poor prognosis in MDS patients, while high ApoA1 level ( OR=0.785, 95% CI: 0.658~0.937, P=0.007) was a protective factor ( P<0.05). Conclusion:The decrease in ApoA1 levels and the increase in CTLA-4 and S100A9 levels in MDS patients are associated with poor prognosis.

Objective:To investigate the relationship between serum apolipoprotein A1 (ApoA1), cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), proinflammatory protein S100A9 (S100A9) and prognosis in patients with myelodysplastic syndrome (MDS).Methods:122 MDS patients visited Qinhuangdao First Hospital from January 2020 to January 2021 were selected as the research objects. After three years of follow up, patients were divided into survival group and death group based on survival status. The differences in ApoA1, CTLA-4, and S100A9 between the death group and the survival group were compared. Measurement data with normal distribution was expressed as " xˉ±s", independent sample t-test was used on comparison between groups. Counting data was expressed as rate or composition ratio, χ2 test was used on comparison between groups. Univariate and multivariate Logistic regression analysis were used to analyze factors related to death. Results:There was no lost to follow up patients after three years of follow up. Among those 122 patients, 92 survived and 30 died. The ratio of bone marrow primitive cells>5%, IPSS-R score, serum CTLA-4, and S100A9 levels in the survival group were (2.89±2.15), (3.13±1.95) points, (5.12±1.59) μg/L, (1643.98±429.65)ng/L, respectively, lower than (5.67±3.76), (5.12±2.36) points, (28.67±6.98) μg/L, (2895.64±553.62) ng/L in the death group ( t=5.03, 4.60, 30.27, 12.87, respectively, all P<0.01). The relative high-risk ratio of IPSS-R stratification in the survival group was 63.04%,(58/92) which was lower than the 86.67%(26/30) in the death group ( χ2=5.89, P=0.015). The absolute values of hemoglobin, lymphocytes and neutrophils, and values of platelets and ApoA1 in the survival group were(86.74±12.69)g/L, (1.41±0.23)×10 9/L, (1.42±0.55)×10 9/L, (59.98±21.37)×10 9/L, (1.09±0.40) g/L respectively, which were higher than (65.58±10.89)g/L, (0.68±0.17)×10 9/L, (0.96±0.31)×10 9/L, (42.85±20.95)×10 9/L, (0.91±0.36)g/L in the death group ( t=8.20, 16.00, 4.35, 7.90, 2.19; respectively, P<0.001, <0.001, <0.001, <0.001, =0.030). Multivariate Logistic regression model analysis showed that, bone marrow blasts cells>5% ( OR=1.732, 95% CI: 1.188~2.523, P=0.004), relatively high IPSS-R stratification ( OR=1.815, 95% CI: 1.332~2.474, P<0.001), high IPSS-R score ( OR=1.785, 95% CI: 1.259~2.529, P=0.001), high CTLA-4 level ( OR=2.156, 95% CI: 1.482~3.134, P<0.001) and high S100A9 level ( OR=1.787, 95% CI: 1.218~2.625, P=0.003) were risk factors for poor prognosis in MDS patients, while high ApoA1 level ( OR=0.785, 95% CI: 0.658~0.937, P=0.007) was a protective factor ( P<0.05). Conclusion:The decrease in ApoA1 levels and the increase in CTLA-4 and S100A9 levels in MDS patients are associated with poor prognosis.

BACKGROUND:Bile acid metabolism plays a crucial role in the development and progression of Crohn's disease.There is no research on changes in bile acid metabolism and key target genes following treatment with biological agents.OBJECTIVE:To investigate the expression characteristics of bile acid metabolism-related genes in patients with Crohn's disease,identify key genes associated with response to biological agents.METHODS:Transcriptome data were obtained through the GEO database to analyze differentially expressed genes between inflammation-control groups and inflammation-treatment groups.GO,KEGG,and GSEA enrichment analyses were used to evaluate the effects of biological agent therapy on bile acid metabolism.Protein-protein interaction network and WGCNA algorithm were employed to analyze differentially expressed genes,identifying modules closely related to biological agent treatment response,which led to the determination of UGT2A3 as a key gene in bile acid metabolism.In the inflammation group of the GSE186582 dataset,samples were divided into high and low expression groups based on UGT2A3 levels to study its relationship with immune infiltration and explore the interaction between UGT2A3 and the immune microenvironment.Clinical characteristics and intestinal manifestations were compared between high and low expression groups,and correlations between UGT2A3 and clinical indicators(C-reactive protein,erythrocyte sedimentation rate,Crohn's disease activity index,and Crohn's disease endoscopic activity score)were investigated.The competing endogenous RNA regulatory network of UGT2A3 was constructed,and its upstream miRNA was functionally enriched to explore the molecular mechanism of UGT2A3 in bile acid metabolism.Single-cell analysis and clustering were performed using high-throughput sequencing data of GSE134809 to observe the expression of UGT2A3 in different samples and cell populations.Colon tissue samples from untreated and biologic-treated Crohn's disease patients and healthy colon tissue samples from patients with intestinal polyps were collected,and UGT2A3 expression was detected by immunohistochemistry,qRT-PCR,and western blot assay.Fresh feces from Crohn's disease patients and healthy controls were collected to detect bile acid levels,and the relationship between UGT2A3 and fecal bile acid levels was analyzed.RESULTS AND CONCLUSION:A total of 11 bile acid metabolism-related genes were screened,showing significant changes in gene expression after biological agent therapy.GO and KEGG enrichment analyses revealed that intestinal nutrient absorption and metabolic processes normalized after treatment,while leukocyte chemotaxis and inflammatory response pathway activity decreased.GSEA analysis revealed significant enrichment of bile acid metabolism-related pathways after treatment.Protein-protein interaction network construction and WGCNA analysis identified UGT2A3 as a key gene closely associated with treatment response.UGT2A3 expression was significantly decreased in inflamed tissues of Crohn's disease patients and returned to normal levels after biological agent therapy.This result was confirmed in clinical specimens.UGT2A3 expression levels showed significant negative correlations with C-reactive protein,erythrocyte sedimentation rate,Crohn's Disease Activity Index,and Crohn's Disease Endoscopic Index of Severity.Receiver Operating Characteristic curve analysis demonstrated that UGT2A3 has good diagnostic value(Area Under Curve AUC=0.801 0)and effectively reflects treatment outcomes.Immune infiltration analysis showed significantly increased infiltration of various immune cells in samples with low UGT2A3 expression,and its expression levels negatively correlated with immune scores,microenvironment scores,and stromal scores.Compared with the low UGT2A3 expression group,patients with high expression showed less fecal occult blood and penetrating inflammation,with milder intestinal strictures and general condition severity.Fecal bile acid analysis revealed that UGT2A3 expression strongly negatively correlated with primary bile acid content and strongly positively correlated with secondary bile acid content.

Objectives:To investigate the expression of signal-induced proliferation-associated 1 (SIPA1) in colorectal cancer tissues and its relationship with patient prognosis. To explore the effects of SIPA1 on proliferation and migration abilities, as well as the possible molecular mechanisms.Methods:Using The Cancer Genome Atlas (TCGA) database to analyze the differential expression of SIPA1 and conduct survival analysis. Then, plotting receiver operating characteristic curve (ROC) and prognosis calibration curve analysis to assess the predictive capability and accuracy of SIPA1 for patient prognosis. Subsequently, verifying the expression levels of SIPA1 in tumor tissues and adjacent normal tissues using immunohistochemistry (IHC) and western blot (WB) assays(from March 1, 2023, to May 1, 2024, pathological specimens of five colorectal cancer patients were selected from the tissue bank of affiliated hospital of Nantong University. tissue microarrays were constructed using both cancerous tissues and adjacent normal tissues), and exploring the correlation between SIPA1 and clinical pathological parameters. Next, establishing SIPA1 stable knockdown cell lines in colorectal cancer cell lines DLD1 and HCT116, and assessing the biological behavior changes of tumor cells after SIPA1 knockdown through cell proliferation, invasion, and migration experiments. Validating the impact of SIPA1 on colorectal cancer cell proliferation in vivo through subcutaneous xenograft experiments in nude mice. Exploring the potential pro-tumor mechanisms of SIPA1 through pathway enrichment analysis, and confirming these using WB experiments. The proliferation, invasion and migration of tumor cells were detected after adding PI3K activator. Lastly, conducting correlation analysis between SIPA1 and immune checkpoint, as well as the association with immune cells in the tumor microenvironment. Results:Differential analysis showed that mRNA expression of SIPA1 in colorectal cancer tissues was significantly higher than that in adjacent normal tissues ( P＜0.05). Prognostic analysis indicated that patients with high expression of SIPA1 had poor overall survival ( P＜0.001), and the expression level of SIPA1was correlated with lymph node invasion ( P＜0.001) and N stage ( P＜0.05). ROC curve and prognosis calibration curve suggest that SIPA1 can effectively predict five-year survival rate of patients (AUC=0.7), and the predictive performance of the model is relatively accurate ( P＜0.001). WB experiments showed a significant increase in the expression level of SIPA1 protein in colorectal cancer specimens ( P＜0.001). Immunohistochemistry results indicated higher staining scores of SIPA1 in tumor tissues. In vitro experiments demonstrated that SIPA1 knockdown significantly inhibited the proliferation, invasion, and migration capabilities of colorectal cancer cells. In DLD1 and HCT116 cells, the SIPA1-knockdown group exhibited significantly lower absorbance compared to the control group (0.89±0.01 vs. 1.57±0.02 and 0.72±0.01 vs. 1.31±0.03, respectively, both P＜0.001). The SIPA1-knockdown group also demonstrated a reduced number of migrated cells relative to the control group (197.93±16.64 vs. 518.48±29.15 and 171.83±12.49 vs. 446.00±21.81, respectively, both P＜0.001). Furthermore, the cell wound-healing rate was significantly lower in the SIPA1-knockdown group than that in the control group [(0.32±0.01)% vs. (0.61±0.01)% and (0.28±0.01)% vs. (0.75±0.01)%, respectively, both P＜0.001]. In vivo animal experiments suggested that SIPA1 knockdown could inhibit tumor growth [(460.35±57.47) mm3 vs (1 177.55±208.24)mm3, (0.76±0.11)g vs (1.43±0.08)g, P＜0.05]. Pathway enrichment analysis revealed significant enrichment of the receptor tyrosine kinase signaling pathway, and SIPA1 knockdown could inhibit the activation of the phosphatidylinositide 3-kinases (PI3K)/protein kinase B (PKB)/glycogen synthase kinase-3β (GSK3β) signaling pathway. The PI3K activator reversed the inhibitory effect of SIPA1 silencing on tumor cell proliferation, invasion and migration. Correlation analysis indicated that high expression of SIPA1 was associated with immune checkpoints and various immunosuppressive cells (all P＜0.05). Conclusions:SIPA1 is highly expressed in colorectal cancer and associated with poor prognosis. SIPA1 may affect the proliferation and migration abilities of tumor cells by regulating the PI3K/AKT/GSK3β signaling pathway.

Objective To investigate the effect and mechanism of osteopontin(OPN)in hepatoma cell migration through galectin-3 binding protein(LGALS3BP).Methods Human hepatoma cell lines SMMC-7721,SMMC-P(stably transfected with empty eukaryotic expression vectors),and SMMC-OPN(stably transfected with the OPN gene)were cultured.mRNA expression levels of OPN and LGALS3BP were measured by RT-qPCR.Western blot assays were used to analyze the relative protein expression of OPN and LGALS3BP and PI3K/AKT pathway.Wound healing assays were performed to explore the cell migration ability.After transfection with LGALS3BP-targeting small interfering RNA(si-LGALS3BP)or negative control small RNA(si-NC)into SMMC-OPN cells,cell migration and relative expression of PI3K/AKT pathway-related proteins were assessed.Results Compared with SMMC-7721 and SMMC-P,the migratory ability of SMMC-OPN cells was significantly reinforced,and expression of LGALS3BP was obviously upregulated at both mRNA and protein levels.Moreover,relative expression of p-PI3K/PI3K and p-AKT/AKT proteins was significantly increased.Wound healing assays showed that the si-LGALS3BP obviously suppressed the migratory ability of SMMC-OPN cells.Furthermore,relative expression of p-PI3K/PI3K and p-AKT/AKT proteins in SMMC-OPN cells was significantly decreased after transfection of si-LGALS3BP.Conclusions OPN activates the PI3K/AKT pathway by upregulating LGALS3BP expression to promote hepatoma cell migration.

Objective:To summarize the clinical and genetic characteristics, treatment and prognosis of four children with Steroid-resistant nephrotic syndrome (SRNS) due to variants of TRPC6 gene. Methods:Clinical data of four children with SRNS admitted to Children′s Hospital Affiliated to Zhengzhou University between May 2020 and August 2022 were collected. Peripheral blood samples were collected from the children and their parents, and whole exome sequencing was carried out. Sanger sequencing was used to verify the pathogenicity of the candidate variants among the children and their parents.Results:All of the four children were found to harbor heterozygous variants of the TRPC6 gene, including c. 523C>T (p.R175W), c. 1327T>A (p.F443I), c. 430G>C (p.E144Q) (unreported previously), and c. 523C>T (p.R175W), which were all missense variants. Two of the children have shown a simple type, whilst two have shown a nephritis type, none had extrarenal phenotype. Comprehensive renal pathology of three children revealed focal segmental glomerulosclerosis (FSGS). Two children were treated with steroids combined with calcineurin inhibitors (CNIs), among whom one showed significant improvement in symptoms. Conclusion:Discoveries of the novel c. 430G>C variant and the new SRNS phenotype of the c. 1327T>A variant have expanded the mutational and phenotypic spectrum of the TRPC6 gene, which has provided a reference for clinical diagnosis and genetic counseling for the families.

Objective To observe the effects of repeated intravitreal injections of ranibizumab and aflibercept on cor-neal morphology of patients with neovascular age-related macular degeneration(nAMD),diabetic macular edema(DME)or retinal vein obstruction(RVO).Methods In this prospective study,64 patients(64 eyes)who underwent therapy in the injection center of the Ophthalmology Department of our hospital from June 2021 to June 2022 were enrolled,including 19 nAMD patients,20 DME patients and 25 RVO patients.Among these patients,29 were treated with aflibercept(40 g·L-1)and 35 were treated with ranibizumab(10 g·L-1).Monocular injections were adopted for all patients,and 3+pro re nata(PRN)therapy was used.Confocal microscope was used for corneal nerve examination,and corneal endo-thelial microscope was used to measure corneal thickness(CT)and corneal endothelial cells.The CT,corneal endothelial cell density(ECD),coefficient of variation(CV),average cell size(ACS),proportion of hexagonal cells(Hex％),cor-neal nerve fiber length(CNFL),corneal nerve fiber density(CNFD)of patients with nAMD,DME and RVO after repeated intravitreal injections of anti-vascular endothelial growth factor(VEGF)drugs were compared,and those parameters at 1 month after injection of different anti-VEGF drugs were compared with the baseline.Results Before injection,ECD in the DME group was lower than that in the nAMD and RVO groups,and the ACS in the DME group was higher than that in the nAMD and RVO groups(all P＜0.05).There was no significant difference in the other indexes among the three groups(all P＞0.05).After 3 injections of anti-VEGF drugs,the ECD in the DME group was lower than that in the nAMD and RVO groups,the ACS in the DME group was higher than that in the nAMD and RVO groups,and the CNFL in the DME group was lower than that in the nAMD and RVO groups(all P＜0.05).The ECD decreased compared with that before injection from the 2nd injection of aflibercept in the nAMD group(all P＜0.05).Hex％decreased significantly after each injection compared with the baseline(all P＜0.05).Other indexes have no significant differences from the baseline(all P＞0.05).In the RVO group,ECD decreased from the 2nd ranibizumab injection compared with the baseline(all P＜0.05).Conclu-sion Repeated intravitreal injections of anti-VEGF drugs can reduce the Hex％and ECD to a certain extent.After injec-tions,CNFL in the DME group is significantly lower than that in the nAMD and RVO groups.